Ligands for monoamine receptors and transporters, and methods of use thereof

ABSTRACT

One aspect of the present invention relates to heterocyclic compounds. A second aspect of the present invention relates to the use of the heterocyclic compounds as ligands for various mammalian cellular receptors, including dopamine, serotonin, or norepinephrine transporters. The compounds of the present invention will find use in the treatment of numerous ailments, conditions and diseases which afflict mammals, including but not limited to addiction, anxiety, depression, sexual dysfunction, hypertension, migraine, Alzheimer&#39;s disease, obesity, emesis, psychosis, schizophrenia, Parkinson&#39;s disease, inflammatory pain, neuropathic pain, Lesche-Nyhane disease, Wilson&#39;s disease, and Tourette&#39;s syndrome. An additional aspect of the present invention relates to the synthesis of combinatorial libraries of the heterocyclic compounds, and the screening of those libraries for biological activity, e.g., in assays based on dopamine transporters.

RELATED APPLICATIONS

This application claims the benefit of priority to: U.S. ProvisionalPatent Application Ser. No. 60/231,667, filed Sep. 11, 2000; U.S.Provisional Patent Application Ser. No. 60/273,530, filed Mar. 5, 2001;and U.S. Provisional Patent Application Ser. No. 60/298,057, filed Jun.13, 2001.

BACKGROUND OF THE INVENTION

Dopamine, norepinephrine and serotonin are mammalian monoamineneurotransmitters that play important roles in a wide variety ofphysiological processes. Therefore, compounds that selectively modulatethe activity of these three neurotransmitters, either individually, inpairs, or as a group, promise to serve as agents effective in thetreatment of a wide range of maladies, conditions and diseases thatafflict mammals due to atypical activities of these neurotransmitters.

For example, depression is believed to result from dysfunction in thenoradrenergic, dopaminergic, or serotonergic systems. Furthermore, thenoradrenergic system appears to be associated with increased drive,whereas the serotonergic system relates more to changes in mood.Therefore, it is possible that the different symptoms of depression maybenefit from drugs acting mainly on one or the other of theseneurotransmitter systems. On the other hand, a single compound thatselectively affects both the noradrenergic and serotonergic systemsshould prove effective in the treatment of depression comprisingsymptoms related to dysfunction in both systems.

Dopamine plays a major role in addiction. Many of the concepts thatapply to dopamine apply to other neurotransmitters as well. As achemical messenger, dopamine is similar to adrenaline. Dopamine affectsbrain processes that control movement, emotional response, and abilityto experience pleasure and pain. Regulation of dopamine plays a crucialrole in our mental and physical health. Neurons containing theneurotransmitter dopamine are clustered in the midbrain in an areacalled the substantia nigra. In Parkinson's disease, thedopamine-transmitting neurons in this area die. As a result, the brainsof people with Parkinson's disease contain almost no dopamine. To helprelieve their symptoms, these patients are given L-DOPA, a drug that canbe converted in the brain to dopamine.

Certain drugs are known as dopamine agonists. These drugs bind todopamine receptors in place of dopamine and directly stimulate thosereceptors. Some dopamine agonists are currently used to treatParkinson's disease. These drugs can stimulate dopamine receptors evenin someone without dopamine-secreting neurons. In contrast to dopamineagonists, dopamine antagonists are drugs that bind but don't stimulatedopamine receptors. Antagonists can prevent or reverse the actions ofdopamine by keeping dopamine from activating receptors.

Dopamine antagonists are traditionally used to treat schizophrenia andrelated mental disorders. A person with schizophrenia may have anoveractive dopamine system. Dopamine antagonists can help regulate thissystem by “turning down” dopamine activity.

Cocaine and other drugs of abuse can alter dopamine function. Such drugsmay have very different actions. The specific action depends on whichdopamine receptors and brain regions the drugs stimulate or block, andhow well the compounds mimic dopamine. Drugs such as cocaine andamphetamine produce their effects by changing the flow ofneurotransmitters. These drugs are defined as indirect acting becausethey depend on the activity of neurons. In contrast, some drugs bypassneurotransmitters altogether and act directly on receptors.

Use of these two types of drugs can lead to very different results intreating the same disease. As mentioned earlier, people with Parkinson'sdisease lose neurons that contain dopamine. To compensate for this loss,the body produces more dopamine receptors on other neurons. Indirectagonists are not very effective in treating the disease since theydepend on the presence of dopamine neurons. In contrast, direct agonistsare more effective because they stimulate dopamine receptors even whendopamine neurons are missing.

Certain drugs increase dopamine concentrations by preventing dopaminereuptake, leaving more dopamine in the synapse. An example is the widelyabused stimulant drug, cocaine. Another example is methylphenidate, usedtherapeutically to treat childhood hyperkinesis and symptoms ofnarcolepsy.

Sensitization or desensitization normally occurs with drug exposure.However, addiction or mental illness can tamper with the reuptakesystem. This disrupts the normal levels of neurotransmitters in thebrain and can lead to faulty desensitization or sensitization. If thishappens in a region of the brain that serves emotion or motivation, theindividual can suffer severe consequences. For example, cocaine preventsdopamine reuptake by binding to proteins that normally transportdopamine. Not only does cocaine “bully” dopamine out of the way, it alsohangs on to the transport proteins much longer than dopamine does. As aresult, more dopamine remains to stimulate neurons, which causes aprolonged feelings of pleasure and excitement. Amphetamine alsoincreases dopamine levels. Again, the result is over-stimulation ofthese pleasure-pathway nerves in the brain.

Dopamine activity is implicated in the reinforcing effects of cocaine,amphetamine and natural rewards. However, dopamine abnormalities arealso believed to underlie some of the core attention deficits seen inacute schizophrenics.

Norepinephrine, also called noradrenaline, is a neurotransmitter thatalso acts as a hormone. As a neurotransmitter, norepinephrine helps toregulate arousal, dreaming, and moods. As a hormone, it acts to increaseblood pressure, constrict blood vessels and increase heartrate-responses that occur when we feel stress.

Serotonin (5-hydroxytryptamine, 5-HT) is widely distributed in animalsand plants, occurring in vertebrates, fruits, nuts, and venoms. A numberof congeners of serotonin are also found in nature and have been shownto possess a variety of peripheral and central nervous systemactivities. Serotonin may be obtained from a variety of dietary sources;however, endogenous 5-HT is synthesized in situ from tryptophan throughthe actions of the enzymes tryptophan hydroxylase and aromatic L-aminoacid decarboxylase. Both dietary and endogenous 5-HT are rapidlymetabolized and inactivated by monoamine oxidase and aldehydedehydrogenase to the major metabolite, 5-hydroxyindoleacetic acid(5-HIAA).

Serotonin is implicated in the etiology or treatment of variousdisorders, particularly those of the central nervous system, includinganxiety, depression, obsessive-compulsive disorder, schizophrenia,stroke, obesity, pain, hypertension, vascular disorders, migraine, andnausea. Recently, understanding of the role of 5-HT in these and otherdisorders has advanced rapidly due to increasing understanding of thephysiological role of various serotonin receptor subtypes.

It is currently estimated that up to 30% of clinically diagnosed casesof depression are resistant to all forms of drug therapy. To achieve aneffective therapy for such patients, it is logical to develop drugs thatpossess reuptake inhibition profiles different from those of drugscurrently available on the market. For example, the exact role ofdopamine in depressive illness is far from clear, however, interventionin the dopamine system may hold promise for the treatment of a subset ofmajor depression.

SUMMARY OF THE INVENTION

One aspect of the present invention relates to heterocyclic compounds. Asecond aspect of the present invention relates to the use of theheterocyclic compounds as ligands for various mammalian cellularreceptors, including dopamine, serotonin, or norepinephrinetransporters.

The compounds of the present invention will find use in the treatment ofnumerous ailments, conditions and diseases which afflict mammals,including but not limited to addiction, anxiety, depression, sexualdysfunction, hypertension, migraine, Alzheimer's disease, obesity,emesis, psychosis, analgesia, schizophrenia, Parkinson's disease,restless leg syndrome, sleeping disorders, attention deficithyperactivity disorder, irritable bowel syndrome, premature ejaculation,menstrual dysphoria syndrome, urinary incontinence, inflammatory pain,neuropathic pain, Lesche-Nyhane disease, Wilson's disease, andTourette's syndrome. An additional aspect of the present inventionrelates to the synthesis of combinatorial libraries of the heterocycliccompounds, and the screening of those libraries for biological activity,e.g., in assays based on dopamine transporters.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts an ORTEP drawing of compound 124, which was the basis forthe assignment of its absolute stereochemistry.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides heterocyclic compounds, and combinatoriallibraries thereof. Furthermore, the present invention providesheterocyclic compounds that are ligands for dopamine, serotonin, ornorepinephrine receptors or transporters, and methods of use thereof forthe treatment of numerous ailments, conditions and diseases whichafflict mammals, including but not limited to addiction, anxiety,depression, sexual dysfunction, hypertension, migraine, Alzheimer'sdisease, obesity, emesis, psychosis, analgesia, schizophrenia,Parkinson's disease, restless leg syndrome, sleeping disorders,attention deficit hyperactivity disorder, irritable bowel syndrome,premature ejaculation, menstrual dysphoria syndrome, urinaryincontinence, inflammatory pain, neuropathic pain, Lesche-Nyhanedisease, Wilson's disease, and Tourette's syndrome. The presentinvention also relates to pharmaceutical formulations of theheterocyclic compounds.

In certain embodiments, compounds of the present invention are ligandsfor mammalian receptors for dopamine, norepinephrine, serotonin, any twoof these three neurotransmitters or all of them. In certain embodiments,compounds of the present invention are ligands for mammaliantransporters of dopamine, norepinephrine, serotonin, any two of thesethree neurotransmitters or all of them. In certain embodiments,compounds of the present invention are agonists of mammalian receptorsfor dopamine, norepinephrine, serotonin, any two of these threeneurotransmitters or all of them. In certain embodiments, compounds ofthe present invention are antagonists or inverse agonists of mammalianreceptors for dopamine, norepinephrine, serotonin, any two of thesethree neurotransmitters or all of them. In certain embodiments,compounds of the present invention are agonists of mammaliantransporters of dopamine, norepinephrine, serotonin, any two of thesethree neurotransmitters or all of them. In certain embodiments,compounds of the present invention are antagonists or inverse agonistsof mammalian transporters of dopamine, norepinephrine, serotonin, anytwo of these three neurotransmitters or all of them.

In certain embodiments, compounds of the present invention are ligandsfor mammalian dopamine receptors. In certain embodiments, compounds ofthe present invention are ligands for mammalian dopamine transporters.In certain embodiments, compounds of the present invention are agonistsof mammalian dopamine receptors. In certain embodiments, compounds ofthe present invention are antagonists or inverse agonists of mammaliandopamine receptors. In certain embodiments, compounds of the presentinvention are agonists of mammalian dopamine transporters. In certainembodiments, compounds of the present invention are antagonists orinverse agonists of mammalian dopamine transporters.

The mammalian dopamine receptor and transporter are members of a familyof cell surface proteins that permit intracellular transduction ofextracellular signals. Cell surface proteins provide eukaryotic andprokaryotic cells a means to detect extracellular signals and transducesuch signals intracellularly in a manner that ultimately results in acellular response or a concerted tissue or organ response. Cell surfaceproteins, by intracellularly transmitting information regarding theextracellular environment via specific intracellular pathways induce anappropriate response to a particular stimulus. The response may beimmediate and transient, slow and sustained, or some mixture thereof. Byvirtue of an array of varied membrane surface proteins, eukaryotic cellsare exquisitely sensitive to their environment.

Extracellular signal molecules, such as growth hormones, vasodilatorsand neurotransmitters, exert their effects, at least in part, viainteraction with cell surface proteins. For example, some extracellularsignal molecules cause changes in transcription of target gene viachanges in the levels of secondary messengers, such as cAMP. Othersignals, indirectly alter gene expression by activating the expressionof genes, such as immediate-early genes that encode regulatory proteins,which in turn activate expression of other genes that encodetranscriptional regulatory proteins. For example, neuron gene expressionis modulated by numerous extracellular signals, includingneurotransmitters and membrane electrical activity. Transsynapticsignals cause rapid responses in neurons that occur over a period oftime ranging from milliseconds, such as the opening of ligand-gatedchannels, to seconds and minutes, such as second messenger-mediatedevents. Genes in neural cells that are responsive to Transsynapticstimulation and membrane electrical activity, include genes, calledimmediate early genes, whose transcription is activated rapidly, withinminutes, and transiently (see, e.g., Sheng et al. (1990) Neuron 4:477-485), and genes whose expression requires protein synthesis andwhose expression is induced or altered over the course of hours.

Cell surface receptors and ion channels are among the cell surfaceproteins that respond to extracellular signals and initiate the eventsthat lead to this varied gene expression and response. Ion channels andcell surface-localized receptors are ubiquitous and physiologicallyimportant cell surface membrane proteins. They play a central role inregulating intracellular levels of various ions and chemicals, many ofwhich are important for cell viability and function.

Cell surface-localized receptors are membrane spanning proteins thatbind extracellular signalling molecules or changes in the extracellularenvironment and transmit the signal via signal transduction pathways toeffect a cellular response. Cell surface receptors bind circulatingsignal polypeptides, such as neurotransmitters, growth factors andhormones, as the initiating step in the induction of numerousintracellular pathways. Receptors are classified on the basis of theparticular type of pathway that is induced. Included among these classesof receptors are those that bind growth factors and have intrinsictyrosine kinase activity, such as the heparin binding growth factor(HBGF) receptors, and those that couple to effector proteins throughguanine nucleotide binding regulatory proteins, which are referred to asG protein coupled receptors and G proteins, respectively.

The G protein transmembrane signaling pathways consist of threeproteins: receptors, G proteins and effectors. G proteins, which are theintermediaries in transmembrane signaling pathways, are heterodimers andconsist of alpha, beta and gamma subunits. Among the members of a familyof G proteins the alpha subunits differ. Functions of G proteins areregulated by the cyclic association of GTP with the alpha subunitfollowed by hydrolysis of GTP to GDP and dissociation of GDP.

G protein coupled receptors are a diverse class of receptors thatmediate signal transduction by binding to G proteins. Signaltransduction is initiated via ligand binding to the cell membranereceptor, which stimulates binding of the receptor to the G protein. Thereceptor G protein interaction releases GDP, which is specifically boundto the G protein, and permits the binding of GTP, which activates the Gprotein. Activated G protein dissociates from the receptor and activatesthe effector protein, which regulates the intracellular levels ofspecific second messengers. Examples of such effector proteins includeadenyl cyclase, guanyl cyclase, phospholipase C, and others.

G protein-coupled receptors, which are glycoproteins, are known to sharecertain structural similarities and homologies (see, e-g., Gilman, A.G., Ann. Rev. Biochem. 56: 615-649 (1987), Strader, C. D. et al. TheFASEB Journal 3: 1825-1832 (1989), Kobilka, B. K., et al. Nature329:75-79 (1985) and Young et al. Cell 45: 711-719 (1986)). Among the Gprotein-coupled receptors that have been identified and cloned are thesubstance P receptor, the angiotensin receptor, the alpha- andbeta-adrenergic receptors and the serotonin receptors. G protein-coupledreceptors share a conserved structural motif. The general and commonstructural features of the G protein-coupled receptors are the existenceof seven hydrophobic stretches of about 20-25 amino acids eachsurrounded by eight hydrophilic regions of variable length. It has beenpostulated that each of the seven hydrophobic regions forms atransmembrane alpha helix and the intervening hydrophilic regions formalternately intracellularly and extracellularly exposed loops. The thirdcytosolic loop between transmembrane domains five and six is theintracellular domain responsible for the interaction with G proteins.

G protein-coupled receptors are known to be inducible. This inducibilitywas originally described in lower eukaryotes. For example, the cAMPreceptor of the cellular slime mold, Dictyostelium, is induced duringdifferentiation (Klein et al., Science 241: 1467-1472 (1988). During theDictyostelium discoideum differentiation pathway, cAMP, induces highlevel expression of its G protein-coupled receptor. This receptortransduces the signal to induce the expression of the other genesinvolved in chemotaxis, which permits multicellular aggregates to align,organize and form stalks (see, Firtel, R. A., et al. Cell 58: 235-239(1989) and Devreotes, P., Science 245: 1054-1058 (1989)).

DEFINITIONS

For convenience, certain terms employed in the specification, examples,and appended claims are collected here.

The term “cell surface proteins” includes molecules that occur on thesurface of cells, interact with the extracellular environment, andtransmit or transduce information regarding the environmentintracellularly.

The term “extracellular signals” includes a molecule or a change in theenvironment that is transduced intracellularly via cell surface proteinsthat interact, directly or indirectly, with the signal. An extracellularsignal is any compound or substance that in some manner specificallyalters the activity of a cell surface protein. Examples of such signalsinclude, but are not limited to, molecules such as acetylcholine, growthfactors, hormones and other mitogenic substances, such as phorbolmistric acetate (PMA), that bind to cell surface receptors and ionchannels and modulate the activity of such receptors and channels.Extracellular signals also includes as yet unidentified substances thatmodulate the activity of a cell surface protein and thereby affectintracellular functions and that are potential pharmacological agentsthat may be used to treat specific diseases by modulating the activityof specific cell surface receptors.

The term “ED₅₀” means the dose of a drug which produces 50% of itsmaximum response or effect. Alternatively, the dose which produces apre-determined response in 50% of test subjects or preparations.

The term “LD₅₀” means the dose of a drug which is lethal in 50% of testsubjects.

The term “therapeutic index” refers to the therapeutic index of a drugdefined as LD₅₀/ED₅₀

The term “structure-activity relationship (SAR)” refers to the way inwhich altering the molecular structure of drugs alters their interactionwith a receptor, enzyme, etc.

The term “agonist” refers to a compound that mimics the action ofnatural transmitter or, when the natural transmitter is not known,causes changes at the receptor complex in the absence of other receptorligands.

The term “antagonist” refers to a compound that binds to a receptorsite, but does not cause any physiological changes unless anotherreceptor ligand is present.

The term “inverse agonist” refers to a compound that binds to aconstitutively active receptor site and reduces its physiologicalfunction.

The term “competitive antagonist” refers to an antagonist, the effectsof which can be overcome by increased concentration of an agonist.

The term “partial agonist” refers to a compound that binds to a receptorsite but does not produce the maximal effect regardless of itsconcentration.

The term “ligand” refers to a compound that binds at the receptor site.

The term “heteroatom” as used herein means an atom of any element otherthan carbon or hydrogen. Preferred heteroatoms are boron, nitrogen,oxygen, phosphorus, sulfur and selenium.

The term “electron-withdrawing group” is recognized in the art, anddenotes the tendency of a substituent to attract valence electrons fromneighboring atoms, i.e., the substituent is electronegative with respectto neighboring atoms. A quantification of the level ofelectron-withdrawing capability is given by the Hammett sigma (aconstant. This well known constant is described in many references, forinstance, J. March, Advanced Organic Chemistry, McGraw Hill BookCompany, New York, (1977 edition) pp. 251-259. The Hammett constantvalues are generally negative for electron donating groups (σ[P]=0.66for NH₂) and positive for electron withdrawing groups (σ[P]=0.78 for anitro group), σ[P] indicating para substitution. Exemplaryelectron-withdrawing groups include nitro, acyl, formyl, sulfonyl,trifluoromethyl, cyano, chloride, and the like. Exemplaryelectron-donating groups include amino, methoxy, and the like.

The term “alkyl” refers to the radical of saturated aliphatic groups,including straight-chain alkyl groups, branched-chain alkyl groups,cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, andcycloalkyl substituted alkyl groups. In preferred embodiments, astraight chain or branched chain alkyl has 30 or fewer carbon atoms inits backbone (e.g., C₁-C₃₀ for straight chain, C₃-C₃₀ for branchedchain), and more preferably 20 or fewer. Likewise, preferred cycloalkylshave from 3-10 carbon atoms in their ring structure, and more preferablyhave 5, 6 or 7 carbons in the ring structure.

Unless the number of carbons is otherwise specified, “lower alkyl” asused herein means an alkyl group, as defined above, but having from oneto ten carbons, more preferably from one to six carbon atoms in itsbackbone structure. Likewise, “lower alkenyl” and “lower alkynyl” havesimilar chain lengths. Preferred alkyl groups are lower alkyls. Inpreferred embodiments, a substituent designated herein as alkyl is alower alkyl.

The term “aralkyl”, as used herein, refers to an alkyl group substitutedwith an aryl group (e.g., an aromatic or heteroaromatic group).

The terms “alkenyl” and “alkynyl” refer to unsaturated aliphatic groupsanalogous in length and possible substitution to the alkyls describedabove, but that contain at least one double or triple bond respectively.

The term “aryl” as used herein includes 5-, 6- and 7-memberedsingle-ring aromatic groups that may include from zero to fourheteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole,oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazineand pyrimidine, and the like. Those aryl groups having heteroatoms inthe ring structure may also be referred to as “aryl heterocycles” or“heteroaromatics.” The aromatic ring can be substituted at one or morering positions with such substituents as described above, for example,halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl,alkoxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate,phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl,sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromatic orheteroaromatic moieties, —CF₃, —CN, or the like. The term “aryl” alsoincludes polycyclic ring systems having two or more cyclic rings inwhich two or more carbons are common to two adjoining rings (the ringsare “fused rings”) wherein at least one of the rings is aromatic, e.g.,the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls,aryls and/or heterocyclyls.

The terms ortho, meta and para apply to 1,2-, 1,3- and 1,4-disubstitutedbenzenes, respectively. For example, the names 1,2-dimethylbenzene andortho-dimethylbenzene are synonymous.

The terms “heterocyclyl” or “heterocyclic group” refer to 3- to10-membered ring structures, more preferably 3- to 7-membered rings,whose ring structures include one to four heteroatoms. Heterocycles canalso be polycycles. Heterocyclyl groups include, for example, thiophene,thianthrene, furan, pyran, isobenzofuran, chromene, xanthene,phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole,pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole,indole, indazole, purine, quinolizine, isoquinoline, quinoline,phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline,pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine,phenanthroline, phenazine, phenarsazine, phenothiazine, furazan,phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine,piperazine, morpholine, lactones, lactams such as azetidinones andpyrrolidinones, sultams, sultones, and the like. The heterocyclic ringcan be substituted at one or more positions with such substituents asdescribed above, as for example, halogen, alkyl, aralkyl, alkenyl,alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio,sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic orheteroaromatic moiety, —CF₃, —CN, or the like.

The terms “polycyclyl” or “polycyclic group” refer to two or more rings(e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/orheterocyclyls) in which two or more carbons are common to two adjoiningrings, e.g., the rings are “fused rings”. Rings that are joined throughnon-adjacent atoms are termed “bridged” rings. Each of the rings of thepolycycle can be substituted with such substituents as described above,as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate,phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl,ketone, aldehyde, ester, a heterocyclyl, an aromatic or heteroaromaticmoiety, —CF₃, —CN, or the like.

As used herein, the term “nitro” means —NO₂; the term “halogen”designates —F, —Cl, —Br or —I; the term “sulfhydryl” means —SH; the term“hydroxyl” means —OH; and the term “sulfonyl” means —SO₂—.

The terms “amine” and “amino” are art-recognized and refer to bothunsubstituted and substituted amines, e.g., a moiety that can berepresented by the general formula:

wherein R₉, R₁₀ and R′₁₀ each independently represent a group permittedby the rules of valence.

The term “acylamino” is art-recognized and refers to a moiety that canbe represented by the general formula:

wherein R₉ represents a group permitted by the rules of valence, andR′₁₁ represents hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl,aralkyl, or heteroaralkyl.

The term “amido” is art recognized as an amino-substituted carbonyl andincludes a moiety that can be represented by the general formula:

wherein R₉, R₁₀ are as defined above. Preferred embodiments of the amidewill not include imides which may be unstable.

The term “alkylthio” refers to an alkyl group, as defined above, havinga sulfur radical attached thereto. In preferred embodiments, the“alkylthio” moiety is represented by one of —S-alkyl, —S-alkenyl,—S-alkynyl, and —S—(CH₂)_(m)—R₈, wherein m is an integer less than orequal to ten, and R₈ is alkyl, cycloalkyl, alkenyl, aryl, or heteroaryl.Representative alkylthio groups include methylthio, ethyl thio, and thelike.

The term “carbonyl” is art recognized and includes such moieties as canbe represented by the general formula:

wherein X is a bond or represents an oxygen or a sulfur, and R₁₁represents a hydrogen, an alkyl, an alkenyl, —(CH₂)_(m)—R₈ or apharmaceutically acceptable salt, R′₁₁ represents a hydrogen, an alkyl,an alkenyl or —(CH₂)_(m)—R₈, where m and R₈ are as defined above. WhereX is an oxygen and R₁₁ or R′₁₁ is not hydrogen, the formula representsan “ester”. Where X is an oxygen, and R₁₁ is as defined above, themoiety is referred to herein as a carboxyl group, and particularly whenR₁₁ is a hydrogen, the formula represents a “carboxylic acid”. Where Xis an oxygen, and R′₁₁ is hydrogen, the formula represents a “formate”.In general, where the oxygen atom of the above formula is replaced bysulfur, the formula represents a “thiolcarbonyl” group. Where X is asulfur and R₁₁ or R′₁₁ is not hydrogen, the formula represents a“thiolester.” Where X is a sulfur and R₁₁ is hydrogen, the formularepresents a “thiolcarboxylic acid.” Where X is a sulfur and R′₁₁ ishydrogen, the formula represents a “thiolformate.” On the other hand,where X is a bond, and R₁₁ is not hydrogen, the above formula representsa “ketone” group. Where X is a bond, and R₁₁ is hydrogen, the aboveformula represents an “aldehyde” group.

The terms “alkoxyl” or “alkoxy” as used herein refers to an alkyl group,as defined above, having an oxygen radical attached thereto.Representative alkoxyl groups include methoxy, ethoxy, propyloxy,tert-butoxy and the like. An “ether” is two hydrocarbons covalentlylinked by an oxygen. Accordingly, the substituent of an alkyl thatrenders that alkyl an ether is or resembles an alkoxyl, such as can berepresented by one of —O-alkyl, —O-alkenyl, —O-alkynyl, —O—(CH₂)_(m)—R₈,where m and R₈ are described above.

The term “sulfonate” is art recognized and includes a moiety that can berepresented by the general formula:

in which R₄₁ is an electron pair, hydrogen, alkyl, cycloalkyl, or aryl.

The terms triflyl, tosyl, mesyl, and nonaflyl are art-recognized andrefer to trifluoromethanesulfonyl, p-toluenesulfonyl, methanesulfonyl,and nonafluorobutanesulfonyl groups, respectively. The terms triflate,tosylate, mesylate, and nonaflate are art-recognized and refer totrifluoromethanesulfonate ester, p-toluenesulfonate ester,methanesulfonate ester, and nonafluorobutanesulfonate ester functionalgroups and molecules that contain said groups, respectively.

The abbreviations Me, Et, Ph, Tf, Nf, Ts, Ms represent methyl, ethyl,phenyl, trifluoromethanesulfonyl, nonafluorobutanesulfonyl,p-toluenesulfonyl and methanesulfonyl, respectively. A morecomprehensive list of the abbreviations utilized by organic chemists ofordinary skill in the art appears in the first issue of each volume ofthe Journal of Organic Chemistry; this list is typically presented in atable entitled Standard List of Abbreviations. The abbreviationscontained in said list, and all abbreviations utilized by organicchemists of ordinary skill in the art are hereby incorporated byreference.

The term “sulfate” is art recognized and includes a moiety that can berepresented by the general formula:

in which R₄₁ is as defined above.

The term “sulfonylamino” is art recognized and includes a moiety thatcan be represented by the general formula:

The term “sulfamoyl” is art-recognized and includes a moiety that can berepresented by the general formula:

The term “sulfonyl”, as used herein, refers to a moiety that can berepresented by the general formula:

in which R₄₄ is selected from the group consisting of hydrogen, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

The term “sulfoxido” as used herein, refers to a moiety that can berepresented by the general formula:

in which R₄₄ is selected from the group consisting of hydrogen, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, aralkyl, or aryl.

A “selenoalkyl” refers to an alkyl group having a substituted selenogroup attached thereto. Exemplary “selenoethers” which may besubstituted on the alkyl are selected from one of —Se-alkyl,—Se-alkenyl, —Se-alkynyl, and —Se—(CH₂)_(m)—R₈, m and R₈ being definedabove.

Analogous substitutions can be made to alkenyl and alkynyl groups toproduce, for example, aminoalkenyls, aminoalkynyls, amidoalkenyls,amidoalkynyls, iminoalkenyls, iminoalkynyls, thioalkenyls, thioalkynyls,carbonyl-substituted alkenyls or alkynyls.

As used herein, the definition of each expression, e.g. alkyl, m, n,etc., when it occurs more than once in any structure, is intended to beindependent of its definition elsewhere in the same structure.

It will be understood that “substitution” or “substituted with” includesthe implicit proviso that such substitution is in accordance withpermitted valence of the substituted atom and the substituent, and thatthe substitution results in a stable compound, e.g., which does notspontaneously undergo transformation such as by rearrangement,cyclization, elimination, etc.

As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, aromatic and nonaromaticsubstituents of organic compounds. Illustrative substituents include,for example, those described herein above. The permissible substituentscan be one or more and the same or different for appropriate organiccompounds. For purposes of this invention, the heteroatoms such asnitrogen may have hydrogen substituents and/or any permissiblesubstituents of organic compounds described herein which satisfy thevalences of the heteroatoms. This invention is not intended to belimited in any manner by the permissible substituents of organiccompounds.

The phrase “protecting group” as used herein means temporarysubstituents which protect a potentially reactive functional group fromundesired chemical transformations. Examples of such protecting groupsinclude esters of carboxylic acids, silyl ethers of alcohols, andacetals and ketals of aldehydes and ketones, respectively. The field ofprotecting group chemistry has been reviewed (Greene, T. W.; Wuts, P. G.M. Protective Groups in Organic Synthesis, 2^(nd) ed.; Wiley: New York,1991).

Certain compounds of the present invention may exist in particulargeometric or stereoisomeric forms. The present invention contemplatesall such compounds, including cis- and trans-isomers, R- andS-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemicmixtures thereof, and other mixtures thereof, as falling within thescope of the invention. Additional asymmetric carbon atoms may bepresent in a substituent such as an alkyl group. All such isomers, aswell as mixtures thereof, are intended to be included in this invention.

If, for instance, a particular enantiomer of a compound of the presentinvention is desired, it may be prepared by asymmetric synthesis, or byderivation with a chiral auxiliary, where the resulting diastereomericmixture is separated and the auxiliary group cleaved to provide the puredesired enantiomers. Alternatively, where the molecule contains a basicfunctional group, such as amino, or an acidic functional group, such ascarboxyl, diastereomeric salts are formed with an appropriateoptically-active acid or base, followed by resolution of thediastereomers thus formed by fractional crystallization orchromatographic means well known in the art, and subsequent recovery ofthe pure enantiomers. Further, mixtures of stereoisomers may be resolvedusing chiral chromatographic means.

Contemplated equivalents of the compounds described above includecompounds which otherwise correspond thereto, and which have the samegeneral properties thereof, wherein one or more simple variations ofsubstituents are made which do not adversely affect the efficacy of thecompound in binding to monoamine transporters. In general, the compoundsof the present invention may be prepared by the methods illustrated inthe general reaction schemes as, for example, described below, or bymodifications thereof, using readily available starting materials,reagents and conventional synthesis procedures. In these reactions, itis also possible to make use of variants which are in themselves known,but are not mentioned here.

For purposes of this invention, the chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover. Alsofor purposes of this invention, the term “hydrocarbon” is contemplatedto include all permissible compounds having at least one hydrogen andone carbon atom. In a broad aspect, the permissible hydrocarbons includeacyclic and cyclic, branched and unbranched, carbocyclic andheterocyclic, aromatic and nonaromatic organic compounds which can besubstituted or unsubstituted.

Compounds of the Invention

In certain embodiments, a compound of the present invention isrepresented by A:

wherein

X represents C(R₃)₂, O, S, SO, SO₂, NR₂, NC(O)₂R₇, NC(O)OR₂, NS(O)₂R₇,or C═O;

Z represents C(R₃)₂, C(O), O, NR, NC(O)OR, S, SO, or SO₂;

m is 1, 2, 3, 4 or 5;

n is 1 or 2;

p is 0, 1, 2, or 3;

y is 0, 1, or 2;

R represents H, alkyl, cycloalkyl, aryl, heteroaryl, aralkyl, orheteroaralkyl;

R₁ represents H, alkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl;

R and R₁ may be connected through a covalent bond;

R₂ represents independently for each occurrence H, alkyl, fluoroalkyl,aryl, heteroaryl, or cycloalkyl;

R₃ represents independently for each occurrence H, alkyl, aryl, OR₂,OC(O)R₂, CH₂OR₂, or CO₂R₂; wherein any two instances of R₃ may beconnected by a covalent tether whose backbone consists of 1, 2, 3, or 4carbon atoms;

R₄ represents independently for each occurrence H, alkyl, cycloalkyl,aryl, heteroaryl, alkenyl, or OR;

R₅ and R₆ are selected independently for each occurrence from the groupconsisting of H, alkyl, (CH₂)_(p)Y, aryl, heteroaryl, F, OR₂, andOC(O)R₂; or an instance of CR₅R₆ taken together is C(O);

R₇ represents alkyl, cycloalkyl, aryl, heteroaryl, aralkyl, orheteroaralkyl;

R₈ and R₉ are selected independently for each occurrence from the groupconsisting of H, alkyl, (CH₂)_(p)Y, aryl, heteroaryl, F, OR₂, andOC(O)R₂; or an instance of CR₈R₉ taken together is C(O);

Y represents independently for each occurrence OR₂, N(R₂)₂, SR₂, S(O)R₂,S(O)₂R₂, or P(O)(OR₂)₂;

any two instances of R₂ may be connected through a covalent bond;

a covalent bond may connect R₄ and an instance of R₅ or R₆;

any two instances of R₅ and R₆ may be connected through a covalent bond;

any two germinal or vicinal instances of R₈ and R₉ may be connectedthrough a covalent bond; and

the stereochemical configuration at any stereocenter of a compoundrepresented by A is R, S, or a mixture of these configurations.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein Z is O or NR.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein m is 2 or 3.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein n is 1.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein y is 1.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein R₁ representsaryl.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein R₃ representsindependently for each occurrence H or alkyl.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein R₄ representscycloalkyl, aryl, or heteroaryl.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein R₅ and R₆ areselected independently for each occurrence from the group consisting ofH, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein R₈ and R₉ areselected independently for each occurrence from the group consisting ofH, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂; and Z is O or NR.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂; Z is O or NR; and m is 2 or 3.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂; Z is O or NR; and n is 1.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂; Z is O or NR; and y is 1.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂; Z is O or NR; m is 2 or 3; n is 1; and y is 1.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂; Z is O or NR₂; m is 2 or 3; n is 1; y is 1; and R₁ is aryl.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂; Z is O or NR; m is 2 or 3; n is 1; y is 1; R₁ is aryl; and R₃ isH or alkyl.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂; Z is O or NR; m is 2 or 3; n is 1; y is 1; R₁ is aryl; R₃ is Hor alkyl; and R₄ is cycloalkyl, aryl, or heteroaryl.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂; Z is O or NR; m is 2 or 3; n is 1; y is 1; R₁ is aryl; R₃ is Hor alkyl; R is cycloalkyl, aryl, or heteroaryl; and R₅ and R₆ areselected independently for each occurrence from the group consisting ofH, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is C(R₃)₂, O,or NR₂; Z is O or NR; m is 2 or 3; n is 1; y is 1; R₁ is aryl; R₃ is Hor alkyl; R is cycloalkyl, aryl, or heteroaryl; R₅ and R₆ are selectedindependently for each occurrence from the group consisting of H, alkyl,OR₂, aryl, heteroaryl, and F; and R₈ and R₉ are selected independentlyfor each occurrence from the group consisting of H, alkyl, OR₂, aryl,heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is CH₂; Z isO; m is 2; n is 1; y is 1; R₁ is 4 trifluoromethylphenyl or3,4-methylenedioxyphenyl; R₃ is H; R₄ is 4-chlorophenyl; R₅ and R₆ areselected independently for each occurrence from the group consisting ofH and alkyl; and R₈ and R₉ are H.

In certain embodiments, the compounds of the present invention arerepresented by A and the attendant definitions, wherein X is CH₂; Z isO; m is 3; n is 1; y is 1; R₁ is 4 trifluoromethylphenyl; R₃ is H; R₄ is4-chlorophenyl; R₅ and R₆ are selected independently for each occurrencefrom the group consisting of H, OH, and alkyl; and R₈ and R₉ are H.

In assays based on mammalian dopamine, serotonin, or norepinephrinereceptors or transporters, certain compounds according to structure Ahave EC₅₀ values less than 1 μM, more preferably less than 100 nM, andmost preferably less than 10 nM.

In assays based on mammalian dopamine receptors or transporters, certaincompounds according to structure A have EC₅₀ values less than 1 μM, morepreferably less than 100 nM, and most preferably less than 10 nM.

In assays based on mammalian dopamine, serotonin, or norepinephrinereceptors or transporters, certain compounds according to structure Ahave IC₅₀ values less than 1 μM, more preferably less than 100 nM, andmost preferably less than 10 nM.

In assays based on mammalian dopamine receptors or transporters, certaincompounds according to structure A have IC₅₀ values less than 1 μM, morepreferably less than 100 nM, and most preferably less than 10 nM.

In certain embodiments, compounds according to structure A are effectivein the treatment of mammals suffering from addiction, anxiety,depression, sexual dysfunction, hypertension, migraine, Alzheimer'sdisease, obesity, emesis, psychosis, analgesia, schizophrenia,Parkinson's disease, restless leg syndrome, sleeping disorders,attention deficit hyperactivity disorder, irritable bowel syndrome,premature ejaculation, menstrual dysphoria syndrome, urinaryincontinence, inflammatory pain, neuropathic pain, Lesche-Nyhanedisease, Wilson's disease, or Tourette's syndrome.

In certain embodiments, a compound of the present invention isrepresented by B:

wherein

Z represents C(R₃)₂, C(O), O, NR, NC(O)OR, S, SO, or SO₂;

m is 1, 2, 3, 4 or 5;

p is 0, 1, 2, or 3;

y is 0, 1 or 2;

R represents H, alkyl, cycloalkyl, aryl, heteroaryl, aralkyl, orheteroaralkyl;

R₁ represents H, alkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl;

R and R₁ may be connected through a covalent bond;

R₂ represents independently for each occurrence H, alkyl, fluoroalkyl,aryl, heteroaryl, or cycloalkyl;

R₃ represents independently for each occurrence H, alkyl, aryl, OR₂,OC(O)R₂, CH₂OR₂, or CO₂R₂;

R₄ represents independently for each occurrence H, alkyl, cycloalkyl,aryl, heteroaryl, alkenyl, or OR;

R₅ and R₆ are selected independently for each occurrence from the groupconsisting of H, alkyl, (CH₂)_(p)Y, aryl, heteroaryl, F, OR₂, andOC(O)R₂; or an instance of CR₅R₆ taken together is C(O);

R₈ and R₉ are selected independently for each occurrence from the groupconsisting of H, alkyl, (CH₂)_(p)Y, aryl, heteroaryl, F, OR₂, andOC(O)R₂; or an instance of CR₈R₉ taken together is C(O);

Y represents independently for each occurrence OR₂, N(R₂)₂, SR₂, S(O)R₂,S(O)₂R₂, or P(O)(OR₂)₂;

any two instances of R₂ may be connected through a covalent bond;

a covalent bond may connect R₄ and an instance of R₅ or R₆;

any two instances of R₅ and R₆ may be connected through a covalent bond;

any two germinal or vicinal instances of R₈ and R₉ may be connectedthrough a covalent bond; and

the stereochemical configuration at any stereocenter of a compoundrepresented by B is R, S, or a mixture of these configurations.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein Z is O or NR.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein m is 3.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein y is 1.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein R₁ representsaryl.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein R₃ representsindependently for each occurrence H or alkyl.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein R₄ representscycloalkyl, aryl, or heteroaryl.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein R₅ and R₆ areselected independently for each occurrence from the group consisting ofH, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein R₈ and R₉ areselected independently for each occurrence from the group consisting ofH, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein Z is O or NR;and m is 3.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein Z is O or NR;and y is 1.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein Z is O or NR, mis 3; and y is 1.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein Z is O or NR; mis 3; y is 1; and R₁ is aryl.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein Z is O or NR; mis 3; y is 1; R₁ is aryl; and R₃ is H or alkyl.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein Z is O or NR; mis 3; y is 1; R₁ is aryl; R₃ is H or alkyl; and R₄ is cycloalkyl, aryl,or heteroaryl.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein Z is O or NR; mis 3; y is 1; R₁ is aryl; R₃ is H or alkyl; R₄ is cycloalkyl, aryl, orheteroaryl; and R₅ and R₆ are selected independently for each occurrencefrom the group consisting of H, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by B and the attendant definitions, wherein Z is O or NR; mis 3; y is 1; R₁ is aryl; R₃ is H or alkyl; R₄ is cycloalkyl, aryl, orheteroaryl; R₅ and R₆ are selected independently for each occurrencefrom the group consisting of H, alkyl, OR₂, aryl, heteroaryl, and F; andR₈ and R₉ are selected independently for each occurrence from the groupconsisting of H, alkyl, OR₂, aryl, heteroaryl, and F.

In assays based on mammalian dopamine, serotonin, or norepinephrinereceptors or transporters, certain compounds according to structure Bhave EC₅₀ values less than 1 μM, more preferably less than 100 nM, andmost preferably less than 10 nM.

In assays based on mammalian dopamine receptors or transporters, certaincompounds according to structure B have EC₅₀ values less than 1 μM, morepreferably less than 100 nM, and most preferably less than 10 nM.

In assays based on mammalian dopamine, serotonin, or norepinephrinereceptors or transporters, certain compounds according to structure Bhave IC₅₀ values less than 1 μM, more preferably less than 100 nM, andmost preferably less than 10 nM.

In assays based on mammalian dopamine receptors or transporters, certaincompounds according to structure B have IC₅₀ values less than 1 μM, morepreferably less than 100 nM, and most preferably less than 10 nM.

In certain embodiments, compounds according to structure B are effectivein the treatment of mammals suffering from addiction, anxiety,depression, sexual dysfunction, hypertension, migraine, Alzheimer'sdisease, obesity, emesis, psychosis, analgesia, schizophrenia,Parkinson's disease, restless leg syndrome, sleeping disorders,attention deficit hyperactivity disorder, irritable bowel syndrome,premature ejaculation, menstrual dysphoria syndrome, urinaryincontinence, inflammatory pain, neuropathic pain, Lesche-Nyhanedisease, Wilson's disease, or Tourette's syndrome.

In certain embodiments, a compound of the present invention isrepresented by C:

wherein

Z represents C(R₃)₂, C(O), O, NR, NC(O)OR, S, SO, or SO₂;

m is 1, 2, 3, 4 or 5;

p is 0, 1, 2, or 3;

y is 0, 1 or 2;

R represents H, alkyl, cycloalkyl, aryl, heteroaryl, aralkyl, orheteroaralkyl;

R₁ represents H, alkyl aryl, heteroaryl, aralkyl, or heteroaralkyl;

R and R₁ may be connected through a covalent bond;

R₂ represents independently for each occurrence H, alkyl, fluoroalkyl,aryl, heteroaryl, or cycloalkyl;

R₃ represents independently for each occurrence H, alkyl, aryl, OR₂,OC(O)R₂, CH₂OR₂, or CO₂R₂;

R₄ represents independently for each occurrence H, alkyl, cycloalkyl,aryl, heteroaryl, alkenyl, or OR;

R₅ and R are selected independently for each occurrence from the groupconsisting of H, alkyl, (CH₂)Y, aryl, heteroaryl, F, OR₂, and OC(O)R₂;or an instance of CR₅R₆ taken together is C(O);

R₈ and R₉ are selected independently for each occurrence from the groupconsisting of H, alkyl, (CH₂)_(p)Y, aryl, heteroaryl, F, OR₂, andOC(O)R₂; or an instance of CR₉R₉ taken together is C(O);

Y represents independently for each occurrence OR₂, N(R₂)₂, SR₂, S(O)R₂,S(O)₂R₂, or P(O)(OR₂)₂;

any two instances of R₂ may be connected through a covalent bond;

a covalent bond may connect R₄ and an instance of R₅ or R₆;

any two instances of R₅ and R₆ may be connected through a covalent bond;

any two germinal or vicinal instances of R₈ and R₉ may be connectedthrough a covalent bond; and

the stereochemical configuration at any stereocenter of a compoundrepresented by C is R or S, or a mixture of these configurations.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein Z is O or NR.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein m is 3.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein y is 1.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein R₁ representsaryl.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein R₃ representsindependently for each occurrence H or alkyl.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein R₄ representscycloalkyl, aryl, or heteroaryl.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein R₅ and R₆ areselected independently for each occurrence from the group consisting ofH, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein R₈ and R₉ areselected independently for each occurrence from the group consisting ofH, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein Z is O or NR;and m is 3.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein Z is O or NR;and y is 1.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein Z is O or NR; mis 3; and y is 1.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein Z is O or NR; mis 3; y is 1; and R₁ is aryl.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein Z is O or NR; mis 3; y is 1; R₁ is aryl; and R₃ is H or alkyl.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein Z is O or NR; mis 3; y is 1; R₁ is aryl; R₃ is H or alkyl; and R₄ is cycloalkyl, aryl,or heteroaryl.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein Z is O or NR; mis 3; y is 1; R₁ is aryl; R₃ is H or alkyl; R₄ is cycloalkyl, aryl, orheteroaryl; and R₅ and R₆ are selected independently for each occurrencefrom the group consisting of H, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by C and the attendant definitions, wherein Z is O or NR; mis 3; y is 1; R₁ is aryl; R₃ is H or alkyl; R₄ is cycloalkyl, aryl, orheteroaryl; R₅ and R₆ are selected independently for each occurrencefrom the group consisting of H, alkyl, OR₂, aryl, heteroaryl, and F; andR₈ and R₉ are selected independently for each occurrence from the groupconsisting of H, alkyl, OR₂, aryl, heteroaryl, and F.

In assays based on mammalian dopamine, serotonin, or norepinephrinereceptors or transporters, certain compounds according to structure Chave EC₅₀ values less than 1 μM, more preferably less than 100 nM, andmost preferably less than 10 nM.

In assays based on mammalian dopamine receptors or transporters, certaincompounds according to structure C have EC₅₀ values less than 1 μM, morepreferably less than 100 nM, and most preferably less than 10 nM.

In assays based on mammalian dopamine, serotonin, or norepinephrinereceptors or transporters, certain compounds according to structure Chave IC₅₀ values less than 1 μM, more preferably less than 100 nM, andmost preferably less than 10 nM.

In assays based on mammalian dopamine receptors or transporters, certaincompounds according to structure C have IC₅₀ values less than 1 μM, morepreferably less than 100 nM, and most preferably less than 10 nM.

In certain embodiments, compounds according to structure C are effectivein the treatment of mammals suffering from addiction, anxiety,depression, sexual dysfunction, hypertension, migraine, Alzheimer'sdisease, obesity, emesis, psychosis, analgesia, schizophrenia,Parkinson's disease, restless leg syndrome, sleeping disorders,attention deficit hyperactivity disorder, irritable bowel syndrome,premature ejaculation, menstrual dysphoria syndrome, urinaryincontinence, inflammatory pain, neuropathic pain, Lesche-Nyhanedisease, Wilson's disease, or Tourette's syndrome.

In certain embodiments, a compound of the present invention isrepresented by D:

wherein

X represents O, S, SO, SO₂, NR₂, NC(O)R₇, NC(O)OR₂, NS(O)₂R₇, or C═O;

Z represents C(R₃)₂, C(O), O, NR, NC(O)OR, S, SO, or SO₂;

m is 1, 2, 3, 4 or S;

p is 0, 1, 2, or 3;

y is 0, 1, or 2;

R represents H, alkyl, cycloalkyl, aryl, heteroaryl, aralkyl, orheteroaralkyl;

R₁ represents H, alkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl;

R and R₁ may be connected through a covalent bond;

R₂ represents independently for each occurrence H, alkyl, fluoroalkyl,aryl, heteroaryl, or cycloalkyl;

R₃ represents independently for each occurrence H, alkyl, aryl, OR₂,OC(O)R₂, CH₂OR₂, or CO₂R₂;

R₄ represents independently for each occurrence H, alkyl, cycloalkyl,aryl, heteroaryl, alkenyl, or OR;

R₅ and R₆ are selected independently for each occurrence from the groupconsisting of H, alkyl, (CH₂)_(p)Y, aryl, heteroaryl, F, OR₂, andOC(O)R₂; or an instance of CR₈R₉ taken together is C(O);

R₇ represents alkyl, cycloalkyl, aryl, heteroaryl, aralkyl, orheteroaralkyl;

R₈ and R₉ are selected independently for each occurrence from the groupconsisting of H, alkyl, (CH₂)_(p)Y, aryl, heteroaryl, F, OR₂, andOC(O)R₂; or an instance of CR₈R₉ taken together is C(O);

Y represents independently for each occurrence OR₂, N(R₂)₂, SR₂, S(O)R₂,S(O)₂R₂, or P(O)(OR₂)₂;

any two instances of R₂ may be connected through a covalent bond;

a covalent bond may connect R₄ and an instance of R₅ or R₆;

any two instances of R₅ and R₆ may be connected through a covalent bond;

any two germinal or vicinal instances of R₈ and R₉ may be connectedthrough a covalent bond; and

the stereochemical configuration at any stereocenter of a compoundrepresented by D is R, S, or a mixture of these configurations.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein X is O or NR₂.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein Z is O or NR.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein m is 3.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein y is 1.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein R₁ representsaryl.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein R₃ representsindependently for each occurrence H or alkyl.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein R₄ representscycloalkyl, aryl, or heteroaryl.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein R₅ and R₆ areselected independently for each occurrence from the group consisting ofH, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein R₈ and R₉ areselected independently for each occurrence from the group consisting ofH, alkyl, OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein X is O or NR₂;and Z is O or NR.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein X is O or NR₂; Zis O or NR; and m is 3.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein X is O or NR₂; Zis O or NR; and y is 1.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein X is O or NR₂; Zis O or NR; m is 3; and y is 1.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein X is O or NR₂; Zis O or NR; m is 3; y is 1; and R₁ is aryl.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein X is O or NR₂; Zis O or NR; m is 3; y is 1; R₁ is aryl; and R₃ is H or alkyl.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein X is O or NR₂; Zis O or NR; m is 3; y is 1; R₁ is aryl; R₃ is H or alkyl; and R₄ iscycloalkyl, aryl, or heteroaryl.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein X is O or NR₂; Zis O or NR; m is 3; y is 1; R₁ is aryl; R₃ is H or alkyl; R₄ iscycloalkyl, aryl, or heteroaryl; and R₅ and R₆ are selectedindependently for each occurrence from the group consisting of H, alkyl,OR₂, aryl, heteroaryl, and F.

In certain embodiments, the compounds of the present invention arerepresented by D and the attendant definitions, wherein X is O or NR₂; Zis O or NR; m is 3; y is 1; R₁ is aryl; R₃ is H or alkyl; R₄ iscycloalkyl, aryl, or heteroaryl; R₅ and R₆ are selected independentlyfor each occurrence from the group consisting of H, alkyl, OR₂, aryl,heteroaryl, and F; and R and R₅ are selected independently for eachoccurrence from the group consisting of H, alkyl, OR₂, aryl, heteroaryl,and F.

In assays based on mammalian dopamine, serotonin, or norepinephrinereceptors or transporters, certain compounds according to structure Dhave EC₅₀ values less than 1 μM, more preferably less than 100 nM, andmost preferably less than 10 nM.

In assays based on mammalian dopamine receptors or transporters, certaincompounds according to structure D have EC₅₀ values less than 1 μM, morepreferably less than 100 nM, and most preferably less than 10 nM.

In assays based on mammalian dopamine, serotonin, or norepinephrinereceptors or transporters, certain compounds according to structure Dhave IC₅₀ values less than 1 μM, more preferably less than 100 nM, andmost preferably less than 10 nM.

In assays based on mammalian dopamine receptors or transporters, certaincompounds according to structure D have IC₅₀ values less than 1 μM, morepreferably less than 100 nM, and most preferably less than 10 nM.

In certain embodiments, compounds according to structure D are effectivein the treatment of mammals suffering from addiction, anxiety,depression, sexual dysfunction, hypertension, migraine, Alzheimer'sdisease, obesity, emesis, psychosis, analgesia, schizophrenia,Parkinson's disease, restless leg syndrome, sleeping disorders,attention deficit hyperactivity disorder, irritable bowel syndrome,premature ejaculation, menstrual dysphoria syndrome, urinaryincontinence, inflammatory pain, neuropathic pain, Lesche-Nyhanedisease, Wilson's disease, or Tourette's syndrome.

In certain embodiments, the present invention relates to a compoundrepresented by any of the structures outlined above, wherein saidcompound is a single stereoisomer.

In certain embodiments, the present invention relates to a formulation,comprising a compound represented by any of the structures outlinedabove; and a pharmaceutically acceptable excipient.

In certain embodiments, the present invention relates to ligands forreceptors or transporters of dopamine, serotonin, or norepinephrine,wherein the ligands are represented by any of the structures outlinedabove, and any of the sets of definitions associated with one of thosestructures. In certain embodiments, the ligands of the present inventionare antagonists or agonists of receptors or transporters of dopamine,serotonin, or norepinephrine. In any event, the ligands of the presentinvention preferably exert their effect on the dopamine, serotonin, ornorepinephrine receptors or transporters at a concentration less thanabout 1 micromolar, more preferably at a concentration less than about100 nanomolar, and most preferably at a concentration less than 10nanomolar.

In certain embodiments, the selectivity of a ligand for dopaminereceptors or transporters renders that ligand an effective therapeuticagent for an acute or chronic ailment, disease or malady. In certainembodiments, the selectivity of a ligand for dopamine receptors ortransporters consists of a binding affinity for dopamine receptors ortransporters at least a factor of ten greater than its binding affinityfor receptors or transporters of other neurotransmitters. In certainembodiments, the selectivity of a ligand for dopamine receptors ortransporters consists of a binding affinity for dopamine receptors ortransporters at least a factor of one hundred greater than its bindingaffinity for receptors or transporters of other neurotransmitters. Incertain embodiments, the selectivity of a ligand for dopamine receptorsor transporters consists of a binding affinity for dopamine receptors ortransporters at least a factor of one thousand greater than its bindingaffinity for receptors or transporters of other neurotransmitters.

The present invention contemplates pharmaceutical formulations of theligands of the present invention. In certain embodiments, thepharmaceutical formulations will comprise ligands of the presentinvention that selectively effect dopamine receptors or transporters,and thereby have a therapeutic effect on an acute or chronic ailment,disease or malady that is at least in part due to biochemical orphysiological processes associated with dopamine receptors ortransporters. The Background of the Invention (see above) teachesexamples of acute or chronic ailments, diseases or maladies that arecaused or exacerbated by biochemical or physiological processesassociated with dopamine receptors or transporters. One of ordinaryskill in the art will be able to accumulate, by reference to thescientific literature, a more comprehensive list of acute or chronicailments, diseases or maladies that are caused or exacerbated bybiochemical or physiological processes associated with dopaminereceptors or transporters. The present invention contemplatespharmaceutical formulations of ligands of the present invention thatwill be of medicinal value against the aforementioned acute or chronicailments, diseases or maladies.

Biochemical Activity at Cellular Receptors, and Assays to Detect thatActivity

Assaying processes are well known in the art in which a reagent is addedto a sample, and measurements of the sample and reagent are made toidentify sample attributes stimulated by the reagent. For example, onesuch assay process concerns determining in a chromogenic assay theamount of an enzyme present in a biological sample or solution. Suchassays are based on the development of a colored product in the reactionsolution. The reaction develops as the enzyme catalyzes the conversionof a colorless chromogenic substrate to a colored product.

Another assay useful in the present invention concerns determining theability of a ligand to bind to a biological receptor utilizing atechnique well known in the art referred to as a radioligand bindingassay. This assay accurately determines the specific binding of aradioligand to a targeted receptor through the delineation of its totaland nonspecific binding components. Total binding is defined as theamount of radioligand that remains following the rapid separation of theradioligand bound in a receptor preparation (cell homogenates orrecombinate receptors) from that which is unbound. The nonspecificbinding component is defined as the amount of radioligand that remainsfollowing separation of the reaction mixture consisting of receptor,radioligand and an excess of unlabeled ligand. Under this condition, theonly radioligand that remains represents that which is bound tocomponents other that receptor. The specific radioligand bound isdetermined by subtracting the nonspecific from total radioactivitybound. For a specific example of radioligand binding assay for μ-opioidreceptor, see Wang, J. B. et al. FEBS Letters 1994, 338, 217.

Assays useful in the present invention concern determining the activityof receptors the activation of which initiates subsequent intracellularevents in which intracellular stores of calcium ions are released foruse as a second messenger. Activation of some G-protein-coupledreceptors stimulates the formation of inositol triphosphate (IP3, aG-protein-coupled receptor second messenger) through phospholipaseC-mediated hydrolysis of phosphatidylinositol, Berridge and Irvine(1984). Nature 312:315-21. IP3 in turn stimulates the release ofintracellular calcium ion stores.

A change in cytoplasmic calcium ion levels caused by release of calciumions from intracellular stores is used to determine G-protein-coupledreceptor function. This is another type of indirect assay. AmongG-protein-coupled receptors are muscarinic acetylcholine receptors(mAChR), adrenergic receptors, sigma receptors, serotonin receptors,dopamine receptors, angiotensin receptors, adenosine receptors,bradykinin receptors, metabotropic excitatory amino acid receptors andthe like. Cells expressing such G-protein-coupled receptors may exhibitincreased cytoplasmic calcium levels as a result of contribution fromboth intracellular stores and via activation of ion channels, in whichcase it may be desirable although not necessary to conduct such assaysin calcium-free buffer, optionally supplemented with a chelating agentsuch as EGTA, to distinguish fluorescence response resulting fromcalcium release from internal stores. Another type of indirect assayinvolves determining the activity of receptors which, when activated,result in a change in the level of intracellular cyclic nucleotides,e.g., cAMP, cGMP. For example, activation of some dopamine, serotonin,metabotropic glutamate receptors and muscarinic acetylcholine receptorsresults in a decrease in the cAMP or cGMP levels of the cytoplasm.

Furthermore, there are cyclic nucleotide-gated ion channels, e.g., rodphotoreceptor cell channels and olfactory neuron channels [see,Altenhofen, W. et al. (1991) Proc. Natl. Acad. Sci. U.S.A. 88:9868-9872and Dhallan et al. (1990) Nature 347:184-187] that are permeable tocations upon activation by binding of cAMP or cGMP. A change incytoplasmic ion levels caused by a change in the amount of cyclicnucleotide activation of photo-receptor or olfactory neuron channels isused to determine function of receptors that cause a change in cAMP orcGMP levels when activated. In cases where activation of the receptorresults in a decrease in cyclic nucleotide levels, it may be preferableto expose the cells to agents that increase intracellular cyclicnucleotide levels, e.g., forskolin, prior to adding areceptor-activating compound to the cells in the assay. Cell for thistype of assay can be made by co-transfection of a host cell with DNAencoding a cyclic nucleotide-gated ion channel and a DNA encoding areceptor (e.g., certain metabotropic glutamate receptors, muscarinicacetylcholine receptors, dopamine receptors, serotonin receptors and thelike, which, when activated, causes a change in cyclic nucleotide levelsin the cytoplasm.

Any cell expressing a receptor protein which is capable, uponactivation, of directly increasing the intracellular concentration ofcalcium, such as by opening gated calcium channels, or indirectlyaffecting the concentration of intracellular calcium as by causinginitiation of a reaction which utilizes Ca<2+> as a second messenger(e.g., G-protein-coupled receptors), may form the basis of an assay.Cells endogenously expressing such receptors or ion channels and cellswhich may be transfected with a suitable vector encoding one or moresuch cell surface proteins are known to those of skill in the art or maybe identified by those of skill in the art. Although essentially anycell which expresses endogenous ion channel and/or receptor activity maybe used, it is preferred to use cells transformed or transfected withheterologous DNAs encoding such ion channels and/or receptors so as toexpress predominantly a single type of ion channel or receptor. Manycells that may be genetically engineered to express a heterologous cellsurface protein are known. Such cells include, but are not limited to,baby hamster kidney (BHK) cells (ATCC No. CCL10), mouse L cells (ATCCNo. CCLL3), DG44 cells [see, Chasin (1986) Cell. Molec. Genet. 12:555]human embryonic kidney (HEK) cells (ATCC No. CRL1573), Chinese hamsterovary (CHO) cells (ATCC Nos. CRL9618, CCL61, CRL9096), PC12 cells (ATCCNo. CRL1721) and COS-7 cells (ATCC No. CRL1651). Preferred cells forheterologous cell surface protein expression are those that can bereadily and efficiently transfected. Preferred cells include HEK 293cells, such as those described in U.S. Pat. No. 5,024,939.

Any compound which is known to activate ion channels or receptors ofinterest may be used to initiate an assay. Choosing an appropriate ionchannel- or receptor-activating reagent depending on the ion channel orreceptor of interest is within the skill of the art. Directdepolarization of the cell membrane to determine calcium channelactivity may be accomplished by adding a potassium salt solution havinga concentration of potassium ions such that the final concentration ofpotassium ions in the cell-containing well is in the range of about50-150 mM (e.g., 50 mM KCl). With respect to ligand-gated receptors andligand-gated ion channels, ligands are known which have affinity for andactivate such receptors. For example, nicotinic acetyloholine receptorsare known to be activated by nicotine or acetylcholine; similarly,muscarinic and acetylcholine receptors may be activated by addition ofmuscarine or carbamylcholine.

Agonist assays may be carried out on cells known to possess ion channelsand/or receptors to determine what effect, if any, a compound has onactivation or potentiation of ion channels or receptors of interest.Agonist assays also may be carried out using a reagent known to possession channel- or receptor-activating capacity to determine whether a cellexpresses the respective functional ion channel or receptor of interest.

Contacting a functional receptor or ion channel with agonist typicallyactivates a transient reaction; and prolonged exposure to an agonist maydesensitize the receptor or ion channel to subsequent activation. Thus,in general, assays for determining ion channel or receptor functionshould be initiated by addition of agonist (i.e., in a reagent solutionused to initiate the reaction). The potency of a compound having agonistactivity is determined by the detected change in some observable in thecells (typically an increase, although activation of certain receptorscauses a decrease) as compared to the level of the observable in eitherthe same cell, or substantially identical cell, which is treatedsubstantially identically except that reagent lacking the agonist (i.e.,control) is added to the well. Where an agonist assay is performed totest whether or not a cell expresses the functional receptor or ionchannel of interest, known agonist is added to test-cell-containingwells and to wells containing control cells (substantially identicalcell that lacks the specific receptors or ion channels) and the levelsof observable are compared. Depending on the assay, cells lacking theion channel and/or receptor of interest should exhibit substantially noincrease in observable in response to the known agonist. A substantiallyidentical cell may be derived from the same cells from which recombinantcells are prepared but which have not been modified by introduction ofheterologous DNA. Alternatively, it may be a cell in which the specificreceptors or ion channels are removed. Any statistically or otherwisesignificant difference in the level of observable indicates that thetest compound has in some manner altered the activity of the specificreceptor or ion channel or that the test cell possesses the specificfunctional receptor or ion channel.

In an example of drug screening assays for identifying compounds whichhave the ability to modulate ion channels or receptors of interest,individual wells (or duplicate wells, etc.) contain a distinct celltype, or distinct recombinant cell line expressing a homogeneouspopulation of a receptor or ion channel of interest, so that thecompound having unidentified activity may be screened to determinewhether it possesses modulatory activity with respect to one or more ofa variety of functional ion channels or receptors. It is alsocontemplated that each of the individual wells may contain the same celltype so that multiple compounds (obtained from different reagent sourcesin the apparatus or contained within different wells) can be screenedand compared for modulating activity with respect to one particularreceptor or ion channel type.

Antagonist assays, including drug screening assays, may be carried outby incubating cells having functional ion channels and/or receptors inthe presence and absence of one or more compounds, added to the solutionbathing the cells in the respective wells of the microtiter plate for anamount of time sufficient (to the extent that the compound has affinityfor the ion channel and/or receptor of interest) for the compound(s) tobind to the receptors and/or ion channels, then activating the ionchannels or receptors by addition of known agonist, and measuring thelevel of observable in the cells as compared to the level of observablein either the same cell, or substantially identical cell, in the absenceof the putative antagonist.

The assays are thus useful for rapidly screening compounds to identifythose that modulate any receptor or ion channel in a cell. Inparticular, assays can be used to test functional ligand-receptor orligand-ion channel interactions for cell receptors includingligand-gated ion channels, voltage-gated ion channels, G-protein-coupledreceptors and growth factor receptors.

Those of ordinary skill in the art will recognize that assays mayencompass measuring a detectable change of a solution as a consequenceof a cellular event which allows a compound, capable of differentialcharacteristics, to change its characteristics in response to thecellular event. By selecting a particular compound which is capable ofdifferential characteristics upon the occurrence of a cellular event,various assays may be performed. For example, assays for determining thecapacity of a compound to induce cell injury or cell death may becarried out by loading the cells with a pH-sensitive fluorescentindicator such as BCECF (Molecular Probes, Inc., Eugene, Oreg. 97402,Catalog #B1150) and measuring cell injury or cell death as a function ofchanging fluorescence over time.

In a further example of useful assays, the function of receptors whoseactivation results in a change in the cyclic nucleotide levels of thecytoplasm may be directly determined in assays of cells that expresssuch receptors and that have been injected with a fluorescent compoundthat changes fluorescence upon binding cAMP. The fluorescent compoundcomprises cAMP-dependent-protein kinase in which the catalytic andregulatory subunits are each labelled with a different fluorescent-dye[Adams et al. (1991) Nature 349:694-697]. When cAMP binds to theregulatory subunits, the fluorescence emission spectrum changes; thischange can be used as an indication of a change in cAMP concentration.

The function of certain neurotransmitter transporters which are presentat the synaptic cleft at the junction between two neurons may bedetermined by the development of fluorescence in the cytoplasm of suchneurons when conjugates of an amine acid and fluorescent indicator(wherein the fluorescent indicator of the conjugate is an acetoxymethylester derivative e.g., 5-(aminoacetamido)fluorescein; Molecular Probes,Catalog #A1363) are transported by the neurotransmitter transporter intothe cytoplasm of the cell where the ester group is cleaved by esteraseactivity and the conjugate becomes fluorescent.

In practicing an assay of this type, a reporter gene construct isinserted into an eukaryotic cell to produce a recombinant cell which haspresent on its surface a cell surface protein of a specific type. Thecell surface receptor may be endogenously expressed or it may beexpressed from a heterologous gene that has been introduced into thecell. Methods for introducing heterologous DNA into eukaryotic cellsare-well known in the art and any such method may be used. In addition,DNA encoding various cell surface proteins is known to those of skill inthe art or it may be cloned by any method known to those of skill in theart.

The recombinant cell is contacted with a test compound and the level ofreporter gene expression is measured. The contacting may be effected inany vehicle and the testing may be by any means using any protocols,such as serial dilution, for assessing specific molecular interactionsknown to those of skill in the art. After contacting the recombinantcell for a sufficient time to effect any interactions, the level of geneexpression is measured. The amount of time to effect such interactionsmay be empirically determined, such as by running a time course andmeasuring the level of transcription as a function of time. The amountof transcription may be measured using any method known to those ofskill in the art to be suitable. For example, specific mRNA expressionmay be detected using Northern blots or specific protein product may beidentified by a characteristic stain. The amount of transcription isthen compared to the amount of transcription in either the same cell inthe absence of the test compound or it may be compared with the amountof transcription in a substantially identical cell that lacks thespecific receptors. A substantially identical cell may be derived fromthe same cells from which the recombinant cell was prepared but whichhad not been modified by introduction of heterologous DNA.Alternatively, it may be a cell in which the specific receptors areremoved. Any statistically or otherwise significant difference in theamount of transcription indicates that the test compound has in somemanner altered the activity of the specific receptor.

If the test compound does not appear to enhance, activate or induce theactivity of the cell surface protein, the assay may be repeated andmodified by the introduction of a step in which the recombinant cell isfirst tested for the ability of a known agonist or activator of thespecific receptor to activate transcription if the transcription isinduced, the test compound is then assayed for its ability to inhibit,block or otherwise affect the activity of the agonist.

The transcription based assay is useful for identifying compounds thatinteract with any cell surface protein whose activity ultimately altersgene expression. In particular, the assays can be used to testfunctional ligand-receptor or ligand-ion channel interactions for anumber of categories of cell surface-localized receptors, including:ligand-gated ion channels and voltage-gated ion channels, and Gprotein-coupled receptors.

Any transfectable cell that can express the desired cell surface proteinin a manner such the protein functions to intracellularly transduce anextracellular signal may be used. The cells may be selected such thatthey endogenously express the cell surface protein or may be geneticallyengineered to do so. Many such cells are known to those of skill in theart. Such cells include, but are not limited to Ltk<−> cells, PC12 cellsand COS-7 cells.

The preparation of cells which express a receptor or ion channel and areporter gene expression construct, and which are useful for testingcompounds to assess their activities, is exemplified in the Examplesprovided herewith by reference to mammalian Ltk<−> and COS-7 cell lines,which express the Type I human muscarinic (HM1) receptor and which aretransformed with either a c-fos promoter-CAT reporter gene expressionconstruct or a c-fos promoter-luciferase reporter gene expressionconstruct.

Any cell surface protein that is known to those of skill in the art orthat may be identified by those of skill in the art may used in theassay. The cell surface protein may endogenously expressed on theselected cell or it may be expressed from cloned DNA. Exemplary cellsurface proteins include, but are not limited to, cell surface receptorsand ion channels. Cell surface receptors include, but are not limitedto, muscarinic receptors (e.g., human M2 (GenBank accession #M16404);rat M3 (GenBank accession #M16407); human M4 (GenBank accession.#M16405); human M5 (Bonner et al. (1988) Neuron 1:403-410); and thelike); neuronal nicotinic acetylcholine receptors (e.g., the alpha 2,alpha 3 and beta 2 subtypes disclosed in U.S. Ser. No. 504,455 (filedApr. 3, 1990), hereby expressly incorporated by reference herein in itsentirety); the rat alpha 2 subunit (Wada et al. (1988) Science240:330-334); the rat alpha 3 subunit (Boulter et al. (1986) Nature319:368-374); the rat alpha 4 subunit (Goldman et al. (1987) cell48:965-973); the rat alpha 5 subunit (Boulter et al. (1990) J. Biol.Chem. 265:4472-4482); the rat beta 2 subunit (Deneris et al. (1988)Neuron 1:45-54); the rat beta 3 subunit (Deneris et al. (1989) J. Biol.Chem. 264: 6268-6272); the rat beta 4 subunit (Duvoisin et al. (1989)Neuron 3:487-496); combinations of the rat alpha subunits, beta subunitsand alpha and beta subunits; GABA receptors (e.g., the bovine alpha 1and beta 1 subunits (Schofield et al. (1987) Nature 328:221-227); thebovine alpha 2 and alpha 3 subunits (Levitan et al. (1988) Nature335:76-79); the gamma-subunit (Pritchett et al. (1989) Nature338:582-585); the beta 2 and beta 3 subunits (Ymer et alo (1989) EMBO J.8:1665-1670); the delta subunit (Shivers, B. D. (1989) Neuron3:327-337); and the like); glutamate receptors (e.g., receptor isolatedfrom rat brain (Hollmann et al. (1989) Nature 342:643-648); and thelike); adrenergic receptors (e.g., human beta 1 (Frielle et al. (1987)Proc. Natl. Acad. Sci. 84:7920-7924); human alpha 2 (Kobilka et al.(1987) Science 238:650-656); hamster beta 2 (Dixon et al. (1986) Nature321:75-79); and the like); dopamine receptors (e.g., human D2 (Stonnannet al. (1990) Molec. Pharm. 37:1-6); rat (Bunzow et al. (1988) Nature336:783-787); and the like); NGF receptors (e.g., human NGF receptors(Johnson et al. (1986) Cell 47:545-554); and the like); serotoninreceptors (e.g., human 5HT1a Kobilka et al. (1987) Nature 329:75-79);rat 5HT2 (Julius et al. (1990) PNAS 87:928-932); rat 5HT1c (Julius etal. (1988) Science 241:558-564); and the like).

Reporter gene constructs are prepared by operatively linking a reportergene with at least one transcriptional regulatory element. If only onetranscriptional regulatory element is included, it must be a regulatablepromoter. At least one of the selected transcriptional regulatoryelements must be indirectly or directly regulated by the activity of theselected cell-surface receptor whereby activity of the receptor can bemonitored via transcription of the reporter genes.

The construct may contain additional transcriptional regulatoryelements, such as a FIRE sequence, or other sequence, that is notnecessarily regulated by the cell surface protein, but is selected forits ability to reduce background level transcription or to amplify thetransduced signal and to thereby increase the sensitivity andreliability of the assay.

Many reporter genes and transcriptional regulatory elements are known tothose of skill in the art and others may be identified or synthesized bymethods known to those of skill in the art.

A reporter gene includes any gene that expresses a detectable geneproduct, which may be RNA or protein. Preferred reporter genes are thosethat are readily detectable. The reporter gene may also be included inthe construct in the form of a fusion gene with a gene that includesdesired transcriptional regulatory sequences or exhibits other desirableproperties.

Examples of reporter genes include, but are not limited to CAT(chloramphenicol acetyl transferase) (Alton and Vapnek (1979), Nature282: 864-869) luciferase, and other enzyme detection systems, such asbeta-galactosidase; firefly luciferase (deWet et al. (1987), Mol. Cell.Biol. 7:725-737); bacterial luciferase (Engebrecht and Silverman (1984),PNAS 1: 4154-4158; Baldwin et al. (1984), Biochemistry 23: 3663-3667);alkaline phosphatase (Toh et al. (1989) Eur. J. Biochem. 182: 231-238,Hall et al. (1983) J. Mol. Appl. Gen. 2: 101).

Transcriptional control elements include, but are not limited to,promoters, enhancers, and repressor and activator binding sites.Suitable transcriptional regulatory elements may be derived from thetranscriptional regulatory regions of genes whose expression is rapidlyinduced, generally within minutes, of contact between the cell surfaceprotein and the effector protein that modulates the activity of the cellsurface protein. Examples of such genes include, but are not limited to,the immediate early genes (see, Sheng et al. (1990) Neuron 4: 477-485),such as c-fos, Immediate early genes are genes that are rapidly inducedupon binding of a ligand to a cell surface protein. The transcriptionalcontrol elements that are preferred for use in the gene constructsinclude transcriptional control elements from immediate early genes,elements derived from other genes that exhibit some or all of thecharacteristics of the immediate early genes, or synthetic elements thatare constructed such that genes in operative linkage therewith exhibitsuch characteristics. The characteristics of preferred genes from whichthe transcriptional control elements are derived include, but are notlimited to, low or undetectable expression in quiescent cells, rapidinduction at the transcriptional level within minutes of extracellularsimulation, induction that is transient and independent of new proteinsynthesis, subsequent shut-off of transcription requires new proteinsynthesis, and mRNAs transcribed from these genes have a shorthalf-life. It is not necessary for all of these properties to bepresent.

Pharmaceutical Compositions/Formulations

In another aspect, the present invention provides pharmaceuticallyacceptable compositions which comprise a therapeutically-effectiveamount of one or more of the compounds described above, formulatedtogether with one or more pharmaceutically acceptable carriers(additives) and/or diluents. As described in detail below, thepharmaceutical compositions of the present invention may be speciallyformulated for administration in solid or liquid form, including thoseadapted for the following: (1) oral administration, for example,drenches (aqueous or non-aqueous solutions or suspensions), tablets,e.g., those targeted for buccal, sublingual, and systemic absorption,boluses, powders, granules, pastes for application to the tongue, hardgelatin capsules, soft gelatin capsules, mouth sprays, syrups,emulsions, micro-emulsions; (2) parenteral administration, for example,by subcutaneous, intramuscular, intravenous or epidural injection as,for example, a sterile solution or suspension, or sustained-releaseformulation; (3) topical application, for example, as a cream, ointment,or a controlled-release patch or spray applied to the skin; (4)intravaginally or intrarectally, for example, as a pessary, cream orfoam; (5) sublingually; (6) ocularly; (7) transdermally; or (8) nasally.

The phrase “therapeutically-effective amount” as used herein means thatamount of a compound, material, or composition comprising a compound ofthe present invention which is effective for producing some desiredtherapeutic effect in at least a sub-population of cells in an animal ata reasonable benefit/risk ratio applicable to any medical treatment.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The phrase “pharmaceutically-acceptable carrier” as used herein means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, manufacturing aid (e.g.,lubricant, talc magnesium, calcium or zinc stearate, or stearic acid),or solvent encapsulating material, involved in carrying or transportingthe subject compound from one organ, or portion of the body, to anotherorgan, or portion of the body. Each carrier must be “acceptable” in thesense of being compatible with the other ingredients of the formulationand not injurious to the patient. Some examples of materials which canserve as pharmaceutically-acceptable carriers include: (1) sugars, suchas lactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) water; (17) isotonic saline; (18) Ringer's solution; (19)ethyl alcohol; (20) pH buffered solutions; (21) polyesters,polycarbonates and/or polyanhydrides; and (22) other non-toxiccompatible substances employed in pharmaceutical formulations.

As set out above, certain embodiments of the present compounds maycontain a basic functional group, such as amino or alkylamino, and are,thus, capable of forming pharmaceutically-acceptable salts withpharmaceutically-acceptable acids. The term “pharmaceutically-acceptablesalts” in this respect, refers to the relatively non-toxic, inorganicand organic acid addition salts of compounds of the present invention.These salts can be prepared in situ in the administration vehicle or thedosage form manufacturing process, or by separately reacting a purifiedcompound of the invention in its free base form with a suitable organicor inorganic acid, and isolating the salt thus formed during subsequentpurification. Representative salts include the hydrobromide,hydrochloride, sulfate, bisulfate, phosphate, besylate, nitrate,acetate, valerate, oleate, palmitate, stearate, laurate, benzoate,lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate,tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, andlaurylsulphonate salts and the like. (See, for example, Berge et al.(1977) “Pharmaceutical Salts”, J. Pharm. Sci. 66:1-19)

The pharmaceutically acceptable salts of the subject compounds includethe conventional nontoxic salts or quaternary ammonium salts of thecompounds, e.g., from non-toxic organic or inorganic acids. For example,such conventional nontoxic salts include those derived from inorganicacids such as hydrochloride, hydrobromic, sulfuric, sulfamic,phosphoric, nitric, and the like; and the salts prepared from organicacids such as acetic, propionic, succinic, glycolic, stearic, lactic,malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic,phenylacetic, glutamic, benzoic, salicyclic, sulfanilic,2-acetoxybenzoic, fumaric, toluenesulfonic, benzenesulfonic,methanesulfonic, ethane disulfonic, oxalic, isothionic, and the like.

In other cases, the compounds of the present invention may contain oneor more acidic functional groups and, thus, are capable of formingpharmaceutically-acceptable salts with pharmaceutically-acceptablebases. The term “pharmaceutically-acceptable salts” in these instancesrefers to the relatively non-toxic, inorganic and organic base additionsalts of compounds of the present invention. These salts can likewise beprepared in situ in the administration vehicle or the dosage formmanufacturing process, or by separately reacting the purified compoundin its free acid form with a suitable base, such as the hydroxide,carbonate or bicarbonate of a pharmaceutically-acceptable metal cation,with ammonia, or with a pharmaceutically-acceptable organic primary,secondary or tertiary amine. Representative alkali or alkaline earthsalts include the lithium, sodium, potassium, calcium, magnesium, andaluminum salts and the like. Representative organic amines useful forthe formation of base addition salts include ethylamine, diethylamine,ethylenediamine, ethanolamine, diethanolamine, piperazine and the like.(See, for example, Berge et al., supra)

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically-acceptable antioxidants include: (1) watersoluble antioxidants, such as ascorbic acid, cysteine hydrochloride,sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like.

Formulations of the present invention include those suitable for oral,nasal, topical (including buccal and sublingual), rectal, vaginal and/orparenteral administration. The formulations may conveniently bepresented in unit dosage form and may be prepared by any methods wellknown in the art of pharmacy. The amount of active ingredient which canbe combined with a carrier material to produce a single dosage form willvary depending upon the host being treated, the particular mode ofadministration. The amount of active ingredient which can be combinedwith a carrier material to produce a single dosage form will generallybe that amount of the compound which produces a therapeutic effect.Generally, out of one hundred percent, this amount will range from about0.1 percent to about ninety-nine percent of active ingredient,preferably from about 5 percent to about 70 percent, most preferablyfrom about 10 percent to about 30 percent.

Methods of preparing these formulations or compositions include the stepof bringing into association a compound of the present invention withthe carrier and, optionally, one or more accessory ingredients. Ingeneral, the formulations are prepared by uniformly and intimatelybringing into association a compound of the present invention withliquid carriers, or finely divided solid carriers, or both, and then, ifnecessary, shaping the product.

Formulations of the invention suitable for oral administration may be inthe form of capsules, cachets, pills, tablets, lozenges (using aflavored basis, usually sucrose and acacia or tragacanth), powders,granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as anelixir or syrup, or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia) and/or as mouth washes and thelike, each containing a predetermined amount of a compound of thepresent invention as an active ingredient. A compound of the presentinvention may also be administered as a bolus, electuary or paste.

In solid dosage forms of the invention for oral administration(capsules, tablets, pills, dragees, powders, granules, trouches and thelike), the active ingredient is mixed with one or morepharmaceutically-acceptable carriers, such as sodium citrate ordicalcium phosphate, and/or any of the following: (1) fillers orextenders, such as starches, lactose, sucrose, glucose, mannitol, and/orsilicic acid; (2) binders, such as, for example, carboxymethylcellulose,alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3)humectants, such as glycerol; (4) disintegrating agents, such asagar-agar, calcium carbonate, potato or tapioca starch, alginic acid,corn starch, certain silicates, and sodium carbonate; (5) solutionretarding agents, such as paraffin; (6) absorption accelerators, such asquaternary ammonium compounds, and surfactants, such as poloxamer andsodium lauryl sulfate; (7) wetting agents, such as, for example, cetylalcohol, glycerol monostearate, and non-ionic surfactants; (8)absorbents, such as kaolin and bentonite clay; (9) lubricants, such atalc, calcium stearate, magnesium stearate, solid polyethylene glycols,sodium lauryl sulfate, zinc stearate, sodium stearate, stearic acid andmixtures thereof; (10) coloring agents; and (11) controlled releaseagents, such as crospovidone or ethyl cellulose. In the case ofcapsules, tablets and pills, the pharmaceutical compositions may alsocomprise buffering agents. Solid compositions of a similar type may alsobe employed as fillers in soft and hard-shelled gelatin capsules usingsuch excipients as lactose or milk sugars, as well as high molecularweight polyethylene glycols and the like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceuticalcompositions of the present invention, such as dragees, capsules, pillsand granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile, other polymer matrices,liposomes and/or microspheres. They may be formulated for rapid release,e.g., freeze-dried. They may be sterilized by, for example, filtrationthrough a bacteria-retaining filter, or by incorporating sterilizingagents in the form of sterile solid compositions which can be dissolvedin sterile water, or some other sterile injectable medium immediatelybefore use. These compositions may also optionally contain opacifyingagents and may be of a composition that they release the activeingredient(s) only, or preferentially, in a certain portion of thegastrointestinal tract, optionally, in a delayed manner. Examples ofembedding compositions which can be used include polymeric substancesand waxes. The active ingredient can also be in micro-encapsulated form,if appropriate, with one or more of the above-described excipients.

Liquid dosage forms for oral administration of the compounds of theinvention include pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activeingredient, the liquid dosage forms may contain inert diluents commonlyused in the art, such as, for example, water or other solvents,solubilizing agents and emulsifiers, such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor and sesame oils),glycerol, tetrahydroluryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Formulations of the pharmaceutical compositions of the invention forrectal or vaginal administration may be presented as a suppository,which may be prepared by mixing one or more compounds of the inventionwith one or more suitable nonirritating excipients or carrierscomprising, for example, cocoa butter, polyethylene glycol, asuppository wax or a salicylate, and which is solid at room temperature,but liquid at body temperature and, therefore, will melt in the rectumor vaginal cavity and release the active compound.

Formulations of the present invention which are suitable for vaginaladministration also include pessaries, tampons, vaginal tablets, creams,gels, pastes, foams or spray formulations containing such carriers asare known in the art to be appropriate.

Dosage forms for the topical or transdermal administration of a compoundof this invention include powders, sprays, ointments, pastes, creams,lotions, gels, solutions, patches and inhalants. The active compound maybe mixed under sterile conditions with a pharmaceutically-acceptablecarrier, and with any preservatives, buffers, or propellants which maybe required.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients, such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a compound of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane.

Transdermal patches have the added advantage of providing controlleddelivery of a compound of the present invention to the body. Such dosageforms can be made by dissolving or dispersing the compound in the propermedium. Absorption enhancers can also be used to increase the flux ofthe compound across the skin. The rate of such flux can be controlled byeither providing a rate controlling membrane or dispersing the compoundin a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention.

Pharmaceutical compositions of this invention suitable for parenteraladministration comprise one or more compounds of the invention incombination with one or more pharmaceutically-acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containsugars, alcohols, antioxidants, buffers, bacteriostats, solutes whichrender the formulation isotonic with the blood of the intended recipientor suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers which may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms upon the subject compounds may be ensuredby the inclusion of various antibacterial and antifungal agents, forexample, paraben, chlorobutanol, phenol sorbic acid, and the like. Itmay also be desirable to include isotonic agents, such as sugars, sodiumchloride, and the like into the compositions. In addition, prolongedabsorption of the injectable pharmaceutical form may be brought about bythe inclusion of agents which delay absorption such as aluminummonostearate and gelatin.

In some cases, in order to prolong the effect of a drug, it is desirableto slow the absorption of the drug from subcutaneous or intramuscularinjection. This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material having poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally-administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Injectable depot forms are made by forming microencapsule matrices ofthe subject compounds in biodegradable polymers such aspolylactide-polyglycolide. Depending on the ratio of drug to polymer,and the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissue.

When the compounds of the present invention are administered aspharmaceuticals, to humans and animals, they can be given per se or as apharmaceutical composition containing, for example, 0.1 to 99% (morepreferably, 10 to 30%) of active ingredient in combination with apharmaceutically acceptable carrier.

The preparations of the present invention may be given orally,parenterally, topically, or rectally. They are of course given in formssuitable for each administration route. For example, they areadministered in tablets or capsule form, by injection, inhalation, eyelotion, ointment, suppository, etc. administration by injection,infusion or inhalation; topical by lotion or ointment; and rectal bysuppositories. Oral administrations are preferred.

The phrases “parenteral administration” and “administered parenterally”as used herein means modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular,subarachnoid, intraspinal and intrasternal injection and infusion.

The phrases “systemic administration,” “administered systemically,”“peripheral adlministration” and “administered peripherally” as usedherein mean the administration of a compound, drug or other materialother than directly into the central nervous system, such that it entersthe patient's system and, thus, is subject to metabolism and other likeprocesses, for example, subcutaneous administration.

These compounds may be administered to humans and other animals fortherapy by any suitable route of administration, including orally,nasally, as by, for example, a spray, rectally, intravaginally,parenterally, intracisternally and topically, as by powders, ointmentsor drops, including buccally and sublingually.

Regardless of the route of administration selected, the compounds of thepresent invention, which may be used in a suitable hydrated form, and/orthe pharmaceutical compositions of the present invention, are formulatedinto pharmaceutically-acceptable dosage forms by conventional methodsknown to those of skill in the art.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient which is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound of the presentinvention employed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion ormetabolism of the particular compound being employed, the rate andextent of absorption, the duration of the treatment, other drugs,compounds and/or materials used in combination with the particularcompound employed, the age, sex, weight, condition, general health andprior medical history of the patient being treated, and like factorswell known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the effective amount of the pharmaceuticalcomposition required. For example, the physician or veterinarian couldstart doses of the compounds of the invention employed in thepharmaceutical composition at levels lower than that required in orderto achieve the desired therapeutic effect and gradually increase thedosage until the desired effect is achieved.

In general, a suitable daily dose of a compound of the invention will bethat amount of the compound which is the lowest dose effective toproduce a therapeutic effect. Such an effective dose will generallydepend upon the factors described above. Generally, oral, intravenous,intracerebroventricular and subcutaneous doses of the compounds of thisinvention for a patient, when used for the indicated effects, will rangefrom about 0.0001 to about 100 mg per kilogram of body weight per day.

If desired, the effective daily dose of the active compound may beadministered as two, three, four, five, six or more sub-dosesadministered separately at appropriate intervals throughout the day,optionally, in unit dosage forms. Preferred dosing is one administrationper day.

While it is possible for a compound of the present invention to beadministered alone, it is preferable to administer the compound as apharmaceutical formulation (composition).

In another aspect, the present invention provides pharmaceuticallyacceptable compositions which comprise a therapeutically-effectiveamount of one or more of the subject compounds, as described above,formulated together with one or more pharmaceutically acceptablecarriers (additives) and/or diluents. As described in detail below, thepharmaceutical compositions of the present invention may be speciallyformulated for administration in solid or liquid form, including thoseadapted for the following: (1) oral administration, for example,drenches (aqueous or non-aqueous solutions or suspensions), tablets,boluses, powders, granules, pastes for application to the tongue; (2)parenteral administration, for example, by subcutaneous, intramuscularor intravenous injection as, for example, a sterile solution orsuspension; (3) topical application, for example, as a cream, ointmentor spray applied to the skin, lungs, or mucous membranes; or (4)intravaginally or intrarectally, for example, as a pessary, cream orfoam; (5) sublingually or buccally; (6) ocularly; (7) transdermally; or(8) nasally.

The compounds according to the invention may be formulated foradministration in any convenient way for use in human or veterinarymedicine, by analogy with other pharmaceuticals.

The term “treatment” is intended to encompass also prophylaxis, therapyand cure.

The patient receiving this treatment is any animal in need, includingprimates, in particular humans, and other mammals such as equines,cattle, swine and sheep; and poultry and pets in general.

The compound of the invention can be administered as such or inadmixtures with pharmaceutically acceptable carriers and can also beadministered in conjunction with antimicrobial agents such aspenicillins, cephalosporins, aminoglycosides and glycopeptides.Conjunctive therapy, thus includes sequential, simultaneous and separateadministration of the active compound in a way that the therapeuticaleffects of the first administered one is not entirely disappeared whenthe subsequent is administered.

The addition of the active compound of the invention to animal feed ispreferably accomplished by preparing an appropriate feed premixcontaining the active compound in an effective amount and incorporatingthe premix into the complete ration.

Alternatively, an intermediate concentrate or feed supplement containingthe active ingredient can be blended into the feed. The way in whichsuch feed premixes and complete rations can be prepared and administeredare described in reference books (such as “Applied Animal Nutrition”,W.H. Freedman and CO., San Francisco, U.S.A., 1969 or “Livestock Feedsand Feeding” 0 and B books, Corvallis, Ore., U.S.A., 1977).

Combinatorial Libraries

The subject reactions readily lend themselves to the creation ofcombinatorial libraries of compounds for the screening ofpharmaceutical, agrochemical or other biological or medically-relatedactivity or material-related qualities. A combinatorial library for thepurposes of the present invention is a mixture of chemically relatedcompounds which may be screened together for a desired property; saidlibraries may be in solution or covalently linked to a solid support.The preparation of many related compounds in a single reaction greatlyreduces and simplifies the number of screening processes which need tobe carried out. Screening for the appropriate biological,pharmaceutical, agrochemical or physical property may be done byconventional methods.

Diversity in a library can be created at a variety of different levels.For instance, the substrate aryl groups used in a combinatorial approachcan be diverse in terms of the core aryl moiety, e.g., a variegation interms of the ring structure, and/or can be varied with respect to theother substituents.

A variety of techniques are available in the art for generatingcombinatorial libraries of small organic molecules. See, for example,Blondelle et al. (1995) Trends Anal. Chem. 14:83; the Affymax U.S. Pat.Nos. 5,359,115 and 5,362,899: the Ellman U.S. Pat. No. 5,288,514: theStill et al. PCT publication WO 94/08051; Chen et al. (1994) JACS116:2661: Kerr et al. (1993) JACS 115:252; PCT publications WO92/10092,WO93/09668 and WO91/07087; and the Lerner et al. PCT publicationWO93/20242). Accordingly, a variety of libraries on the order of about16 to 1,000,000 or more diversomers can be synthesized and screened fora particular activity or property.

In an exemplary embodiment, a library of substituted diversomers can besynthesized using the subject reactions adapted to the techniquesdescribed in the Still et al. PCT publication WO 94/08051, e.g., beinglinked to a polymer bead by a hydrolyzable or photolyzable group, e.g.,located at one of the positions of substrate. According to the Still etal. technique, the library is synthesized on a set of beads, each beadincluding a set of tags identifying the particular diversomer on thatbead. In one embodiment, which is particularly suitable for discoveringenzyme inhibitors, the beads can be dispersed on the surface of apermeable membrane, and the diversomers released from the beads by lysisof the bead linker. The diversomer from each bead will diffuse acrossthe membrane to an assay zone, where it will interact with an enzymeassay. Detailed descriptions of a number of combinatorial methodologiesare provided below.

A. Direct Characterization

A growing trend in the field of combinatorial chemistry is to exploitthe sensitivity of techniques such as mass spectrometry (MS), e.g.,which can be used to characterize sub-femtomolar amounts of a compound,and to directly determine the chemical constitution of a compoundselected from a combinatorial library. For instance, where the libraryis provided on an insoluble support matrix, discrete populations ofcompounds can be first released from the support and characterized byMS. In other embodiments, as part of the MS sample preparationtechnique, such MS techniques as MALDI can be used to release a compoundfrom the matrix, particularly where a labile bond is used originally totether the compound to the matrix. For instance, a bead selected from alibrary can be irradiated in a MALDI step in order to release thediversomer from the matrix, and ionize the diversomer for MS analysis.

B) Multi in Synthesis

The libraries of the subject method can take the multipin libraryformat. Briefly, Geysen and co-workers (Geysen et al. (1984) PNAS81:3998-4002) introduced a method for generating compound libraries by aparallel synthesis on polyacrylic acid-grated polyethylene pins arrayedin the microtitre plate format. The Geysen technique can be used tosynthesize and screen thousands of compounds per week using the multipinmethod, and the tethered compounds may be reused in many assays.Appropriate linker moieties can also been appended to the pins so thatthe compounds may be cleaved from the supports after synthesis forassessment of purity and further evaluation (c.f., Bray et al. (1990)Tetrahedron Lett 31:5811-5814; Valerio et al. (1991) Anal Biochem197:168-177; Bray et al. (1991) Tetrahedron Lett 32:6163-6166).

C) Divide-Couple-Recombine

In yet another embodiment, a variegated library of compounds can beprovided on a set of beads utilizing the strategy ofdivide-couple-recombine (see, e.g., Houghten (1985) PNAS 82:5131-5135;and U.S. Pat. Nos. 4,631,211; 5,440,016; 5,480,971). Briefly, as thename implies, at each synthesis step where degeneracy is introduced intothe library, the beads are divided into separate groups equal to thenumber of different substituents to be added at a particular position inthe library, the different substituents coupled in separate reactions,and the beads recombined into one pool for the next iteration.

In one embodiment, the divide-couple-recombine strategy can be carriedout using an analogous approach to the so-called “tea bag” method firstdeveloped by Houghten, where compound synthesis occurs on resin sealedinside porous polypropylene bags (Houghten et al. (1986) PNAS82:5131-5135). Substituents are coupled to the compound-bearing resinsby placing the bags in appropriate reaction solutions, while all commonsteps such as resin washing and deprotection are performedsimultaneously in one reaction vessel. At the end of the synthesis, eachbag contains a single compound.

D) Combinatorial Libraries by Light-Directed, Spatially AddressableParallel Chemical Synthesis

A scheme of combinatorial synthesis in which the identity of a compoundis given by its locations on a synthesis substrate is termed aspatially-addressable synthesis. In one embodiment, the combinatorialprocess is carried out by controlling the addition of a chemical reagentto specific locations on a solid support (Dower et al. (1991) Annu RepMed Chem 26:271-280; Fodor, S. P. A. (1991) Science 251:767; Pirrung etal. (1992) U.S. Pat. No. 5,143,854; Jacobs et al. (1994) TrendsBiotechnol 12:19-26). The spatial resolution of photolithography affordsminiaturization. This technique can be carried out through the useprotection/deprotection reactions with photolabile protecting groups.

The key points of this technology are illustrated in Gallop et al.(1994) J Med Chem 37:1233-1251. A synthesis substrate is prepared forcoupling through the covalent attachment of photolabilenitroveratryloxycarbonyl (NVOC) protected amino linkers or otherphotolabile linkers. Light is used to selectively activate a specifiedregion of the synthesis support for coupling. Removal of the photolabileprotecting groups by light (deprotection) results in activation ofselected areas. After activation, the first of a set of amino acidanalogs, each bearing a photolabile protecting group on the aminoterminus, is exposed to the entire surface. Coupling only occurs inregions that were addressed by light in the preceding step. The reactionis stopped, the plates washed, and the substrate is again illuminatedthrough a second mask, activating a different region for reaction with asecond protected building block. The pattern of masks and the sequenceof reactants define the products and their locations. Since this processutilizes photolithography techniques, the number of compounds that canbe synthesized is limited only by the number of synthesis sites that canbe addressed with appropriate resolution. The position of each compoundis precisely known; hence, its interactions with other molecules can bedirectly assessed.

In a light-directed chemical synthesis, the products depend on thepattern of illumination and on the order of addition of reactants. Byvarying the lithographic patterns, many different sets of test compoundscan be synthesized simultaneously; this characteristic leads to thegeneration of many different masking strategies.

E) Encoded Combinatorial Libraries

In yet another embodiment, the subject method utilizes a compoundlibrary provided with an encoded tagging system. A recent improvement inthe identification of active compounds from combinatorial librariesemploys chemical indexing systems using tags that uniquely encode thereaction steps a given bead has undergone and, by inference, thestructure it carries. Conceptually, this approach mimics phage displaylibraries, where activity derives from expressed peptides, but thestructures of the active peptides are deduced from the correspondinggenomic DNA sequence. The first encoding of synthetic combinatoriallibraries employed DNA as the code. A variety of other forms of encodinghave been reported, including encoding with sequenceable bio-oligomers(e.g., oligonucleotides and peptides), and binary encoding withadditional non-sequenceable tags.

1) Tagging with Sequenceable Bio-Oligomers

The principle of using oligonucleotides to encode combinatorialsynthetic libraries was described in 1992 (Brenner et al. (1992) PNAS89:5381-5383), and an example of such a library appeared the followingyear (Needles et al. (1993) PNAS 90:10700-10704). A combinatoriallibrary of nominally 7⁷ (=823,543) peptides composed of all combinationsof Arg, Gln, Phe, Lys, Val, D-Val and Thr (three-letter amino acidcode), each of which was encoded by a specific dinucleotide (TA, TC, CT,AT, TT, CA and AC, respectively), was prepared by a series ofalternating rounds of peptide and oligonucleotide synthesis on solidsupport. In this work, the amine linking functionality on the bead wasspecifically differentiated toward peptide or oligonucleotide synthesisby simultaneously preincubating the beads with reagents that generateprotected OH groups for oligonucleotide synthesis and protected NH₂groups for peptide synthesis (here, in a ratio of 1:20). When complete,the tags each consisted of 69-mers, 14 units of which carried the code.The bead-bound library was incubated with a fluorescently labeledantibody, and beads containing bound antibody that fluoresced stronglywere harvested by fluorescence-activated cell sorting (FACS). The DNAtags were amplified by PCR and sequenced, and the predicted peptideswere synthesized. Following such techniques, compound libraries can bederived for use in the subject method, where the oligonucleotidesequence of the tag identifies the sequential combinatorial reactionsthat a particular bead underwent, and therefore provides the identity ofthe compound on the bead.

The use of oligonucleotide tags permits exquisitely sensitive taganalysis. Even so, the method requires careful choice of orthogonal setsof protecting groups required for alternating co-synthesis of the tagand the library member. Furthermore, the chemical liability of the tag,particularly the phosphate and sugar anomeric linkages, may limit thechoice of reagents and conditions that can be employed for the synthesisof non-oligomeric libraries. In preferred embodiments, the librariesemploy linkers permitting selective detachment of the test compoundlibrary member for assay.

Peptides have also been employed as tagging molecules for combinatoriallibraries. Two exemplary approaches are described in the art, both ofwhich employ branched linkers to solid phase upon which coding andligand strands are alternately elaborated. In the first approach (Kerr JM et al. (1993) J Am Chem Soc 115:2529-2531), orthogonality in synthesisis achieved by employing acid-labile protection for the coding strandand base-labile protection for the compound strand.

In an alternative approach (Nikolaiev et al. (1993) Pept Res 6:161-170),branched linkers are employed so that the coding unit and the testcompound can both be attached to the same functional group on the resin.In one embodiment, a cleavable linker can be placed between the branchpoint and the bead so that cleavage releases a molecule containing bothcode and the compound (Ptek et al. (1991) Tetrahedron Lett32:3891-3894). In another embodiment, the cleavable linker can be placedso that the test compound can be selectively separated from the bead,leaving the code behind. This last construct is particularly valuablebecause it permits screening of the test compound without potentialinterference of the coding groups. Examples in the art of independentcleavage and sequencing of peptide library members and theircorresponding tags has confirmed that the tags can accurately predictthe peptide structure.

2) Non-Sequenceable Tagging: Binary Encoding

An alternative form of encoding the test compound library employs a setof non-sequencable electrophoric tagging molecules that are used as abinary code (Ohlmeyer et al. (1993) PNAS 90:10922-10926). Exemplary tagsare haloaromatic alkyl ethers that are detectable as theirtrimethylsilyl ethers at less than femtomolar levels by electron capturegas chromatography (ECGC). Variations in the length of the alkyl chain,as well as the nature and position of the aromatic halide substituents,permit the synthesis of at least 40 such tags, which in principle canencode 2⁴⁰ (e.g., upwards of 10¹²) different molecules. In the originalreport (Ohlmeyer et al., supra) the tags were bound to about 1% of theavailable amine groups of a peptide library via a photocleavableo-nitrobenzyl linker. This approach is convenient when preparingcombinatorial libraries of peptide-like or other amine-containingmolecules. A more versatile system has, however, been developed thatpermits encoding of essentially any combinatorial library. Here, thecompound would be attached to the solid support via the photocleavablelinker and the tag is attached through a catechol ether linker viacarbene insertion into the bead matrix (Nestler et al. (1994) J Org Chem59:4723-4724). This orthogonal attachment strategy permits the selectivedetachment of library members for assay in solution and subsequentdecoding by ECGC after oxidative detachment of the tag sets.

Although several amide-linked libraries in the art employ binaryencoding with the electrophoric tags attached to amine groups, attachingthese tags directly to the bead matrix provides far greater versatilityin the structures that can be prepared in encoded combinatoriallibraries. Attached in this way, the tags and their linker are nearly asunreactive as the bead matrix itself. Two binary-encoded combinatoriallibraries have been reported where the electrophoric tags are attacheddirectly to the solid phase (Ohlmeyer et al. (1995) PNAS 92:6027-6031)and provide guidance for generating the subject compound library. Bothlibraries were constructed using an orthogonal attachment strategy inwhich the library member was linked to the solid support by aphotolabile linker and the tags were attached through a linker cleavableonly by vigorous oxidation. Because the library members can berepetitively partially photoeluted from the solid support, librarymembers can be utilized in multiple assays. Successive photoelution alsopermits a very high throughput iterative screening strategy: first,multiple beads are placed in 96-well microtiter plates; second,compounds are partially detached and transferred to assay plates; third,a metal binding assay identifies the active wells; fourth, thecorresponding beads are rearrayed singly into new microfiter plates;fifth, single active compounds are identified; and sixth, the structuresare decoded.

EXEMPLIFICATION

The invention now being generally described, it will be more readilyunderstood by reference to the following examples, which are includedmerely for purposes of illustration of certain aspects and embodimentsof the present invention, and are not intended to limit the invention.

Example 1 Synthesis of [1-(4-chloro-phenyl)-cyclobutyl]-(3-hydroxymethylpiperidin-1-yl)-methadone

To a stirred solution of piperidin-3-yl methanol (5.0 g, 0.043 moles)and 1-(4-chloro-phenyl)-cyclobutane carboxylic acid (13.58 g, 0.065moles) in anhydrous dichloromethane (100 mL) was added di-isopropylethyl amine (22.47 mL, 0.219 moles) dropwise. After completion ofaddition solid PyBroP (30.07 g, 0.065 moles) was added to the stirringreaction mixture. The reaction mixture continued stirring at RT for 10 hand was quenched with 10% KOH (aq.). The aqueous layer was extractedwith EtOAc (3×200 mL). The combined organic layers were dried overNa₂SO₄ and concentrated to yield an oil. This crude material waspurified using silica gel chromatography (4:1 hexane:EtOAc-1:1 hexaneEtOAc) to yield 1 as a brown oil (7 g, 0.028 moles, 53%). ¹H(CDCl₃) δ7.26 (4H, s), 4.45 (2H, d, J=12.3 Hz), 3.93 (4H, m), 3.4-1.03 (11H, m).LRMS: M+308.

Example 2 Synthesis of [1-(4-chloro-phenyl)-cyclobutyl]-(3-hydroxymethylpiperidin-1yl)-methanol

A solution of amide 1 (300 mg, 0.977 mmoles) dissolved in anhydroustoluene (10 mL) was cooled to 0° C. RedAl (691 mg, 3.4 mmoles) was addeddropwise to the cooled stirring reaction mixture. After completion ofaddition the reaction continued stirring at RT. After 12 h, 10% KOH wasadded to the reaction mixture. The aqueous layer was extracted withEtOAc (3×5 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated to yield an oil. The crude material was purified usingsilica gel chromatography (4:1 hexanes:EtOAc-4:1 EtOAc:hexanes) to yield2 (230 mg, 0.785 mmole, 80%). ¹H(CDCl₃) δ 7.32-7.12 (4H, m), 3.66-3.47(2H, m), 2.80-1.28 (17H, m). LRMS: M+293.

Example 3 Synthesis of [1-(4-chloro-phenyl)-cyclobutyl]-(3-phenoxymethylpiperidin-1yl)-methadone

A solution of 1 (2.07 g, 6.72 mmoles), triphenylphosphine (2.64 g, 10.08mmole), and phenol (1.27 g, 13.44 mmoles) dissolved in anhydrous ether(50 mL) was cooled in a brine bath to −5° C. DEAD (1.60 mL, 10.08mmoles) dissolved in ether (10 mL) was added to the cooled stirringreaction mixture. After completion of addition, the reaction mixturecontinued stirring at −5° C. After 4 h, the reaction mixture wasconcentrated and crude material was dissolved in a hexane/ethyl acetatemixture (70% hexanes:30% ethyl acetate, 30 mL). Phosphine by-productsprecipitated and were filtered off. The filtrate was concentrated toyield an oil. This oil was purified using silica gel chromatography(100% hexanes-1:1 hexanes: EtOAc) to yield the amine 3 (840 mg, 2.19mmole, 32%). LRMS: M+384.

Example 4 Synthesis of[1-(4-chloro-phenyl)-cyclobutyl]-3-phenoxymethyl-piperidine

A solution of amide 3 (0.5 g, 1.3 mmole) dissolved in anhydrous toluene(15 mL) was cooled to 0° C. RedAl (920 mg, 4.55 mmole) was addeddropwise to the cooled stirring reaction mixture. After completion ofaddition, the reaction continued stirring at RT. After 12 h, 10% KOH wasadded to the reaction mixture. The aqueous layer was extracted withEtOAc (3×20 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated to yield an oil. The crude material was purified usingsilica gel chromatography (100% hexanes-9:1 hexanes:EtOAc) to yield 4(300 mg, 0.811 mmoles, 62%). Enantiomers 5 and 6 were isolated on achiral AD column (100% MeOH). ¹H(CDCl₃) δ 7.35-6.86 (9H, m), 3.80-3.64(2H, m), 2.65-1.15 (15H, m). ¹³C(CDCl₃) δ 159.3, 148.6, 131.0, 129.7,128.0, 127.8, 120.7, 114.7, 70.7, 69.0, 59.2, 56.6, 47.3, 36.4, 31.9,31.8, 27.0, 45.0, 16.3. LRMS: M+370.

Example 5 Synthesis of[1-(4-chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-piperidine

A solution of 2 (50 mg, 0.170 mmoles), triphenylphosphine (66.88 g,0.340 mmoles), and phenol (55.12 mg, 0.340 mmoles) dissolved inanhydrous ether (1.0 mL) was cooled in a brine bath to −5° C. DEAD(40.14 μL, 0.255 mmoles) dissolved in ether (0.5 mL) was added to thecooled stirring reaction mixture. After completion of addition, thereaction mixture continued stirring at −5° C. After 4 h, the reactionmixture was concentrated and crude material was dissolved in ahexane/ethyl acetate mixture (70% hexanes:30% ethyl acetate, 30 mL).Phosphine by-products precipitated and were filtered off. Filtrate wasconcentrated to yield an oil. This oil was purified using silica gelchromatography (4:1 hexanes:EtOAc) to yield 7 (42 mg, 0.096 mmoles,56%). LRMS: M+438.

Example 6 Synthesis of1-[2-(4-Chloro-phenyl)-ethyl]-3-(4-trifluoromethyl-phenoxymethyl)-piperidine

Synthesis of[1-(4-chloro-phenyl)-1-(3-hydroxymethylpiperidin-1yl)-ethanone

To a stirred solution of piperidin-3-yl methanol (1.0 g, 8.7 mmoles) and4-chloro-phenyl)-acetic acid (2.22 g, 13.0 mmoles) in anhydrousdichloromethane (20 mL) was added di-isopropyl ethyl amine (4.55 mL,26.0 mmoles) dropwise. After completion of addition, solid PyBroP (6.06g, 13.0 moles) was added to the stirring reaction mixture. The reactionmixture continued stirring at RT for 10 h and was quenched with 10% KOH(aq.). The aqueous layer was extracted with EtOAc (3×20 mL). Thecombined organic layers were dried over Na₂SO₄ and concentrated to yieldan oil. This crude material was purified using silica gel chromatography(4:1 hexane:EtOAc-95:5 hexane EtOAc) to yield 9 as an oil (2.47 g, 0.028moles, 9.25 mmole, 93%). LRMS: M+267.

Synthesis of2-(4-Chloro-phenyl)-O-[3-(4-trifluoromethyl-phenoxymethyl)-piperidine-1-yl]-ethanone

A solution of 9 (333 mg, 1.24 mmoles), triphenylphosphine (483 mg, 1.86mmoles), and phenol (403 mg, 2.49 mmoles) dissolved in anhydrous ether(4.5 mL) was cooled in a brine bath to −5° C. DEAD (325 mg, 1.86 mmoles)dissolved in ether (0.5 mL) was added to the cooled stirring reactionmixture. After completion of addition, the reaction mixture continuedstirring at −5° C. After 4 h, the reaction mixture was concentrated andcrude material was dissolved in a hexane/ethyl acetate mixture (70%hexanes:30% ethyl acetate, 30 mL). Phosphine by-products precipitatedand were filtered off. Filtrate was concentrated to yield an oil. Thiscrude material was purified using silica gel chromatography (100%hexanes-100% EtOAc) to yield 10 as an oil (249 mg, 0.606 mmoles, 49%).LRMS: M+411.

Compound 10 could be separated into enantiomers 66 and 67 using a chiralAD column (75:25 MeOH:acetonitrile).

Synthesis of1-[2-(4-Chloro-Phenyl)-Ethyl]-3-(4-Trifluoromethyl-Phenoxymethyl)-Piperidine

A solution of amide 10 (100 mg, 0.243 mmole) dissolved in anhydroustoluene (5 mL) was cooled to 0° C. RedAl (172 mg, 0.852 mmole) was addeddropwise to the cooled stirring reaction mixture. After completion ofaddition, the reaction continued stirring at RT. After 12 h, 10% KOH wasadded to the reaction mixture. The aqueous layer was extracted withEtOAc (3×2 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated to yield an oil. The crude material was purified using asilica gel prep plate (100% hexanes-9:1 hexanes:EtOAc) to yield 8 (42mg, 0.106 mmole, 43%). LRMS: M+396.

Example 7 Synthesis of4-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-[2-(4-trifluoromethyl-phenoxymethyl)-[1,4]oxazepane

Synthesis of 3-benzylamino-propan-1-ol

To a stirring solution of benzaldehyde (19.15 mL, 188.3 mmole) dissolvedin anhydrous MeOH (375 mL) was added 3-amino-1-propanol (15.13 mL, 197.8mmole) dropwise. After completion of addition, the reaction mixture washeated to 75° C. After 1 h, the reaction mixture was cooled down to RTand placed in an ice bath. Solid NaBH₄ was added over 20 min. Aftercompletion of addition, the reaction mixture continued stirring at RT.After 10 h, the reaction mixture was concentrated and the white crudematerial was taken up in dichloromethane (300 mL). The organic layer wasextracted with water (200 mL). The aqueous layer was acidified with 10%HCl and then extracted with dichloromethane (3×200 μL). The combinedorganic layers were dried over Na₂SO₄ and concentrated to yield 12 as ayellow oil (23 g, 0.14 moles, 70%). ¹H (CD₃OD) δ 7.40-7.20 (5H, m), 4.98(2H, s), 3.75 (2H, s), 3.64 (1H, t, J=6.2 Hz), 2.70 (2H, t, J=7.1 Hz),1.77 (2H, m). LRMS: M+165.

Synthesis of 4-benzyl-2-chloromethyl-[1,4]oxazepane

A solution of alcohol 12 (3.0 g, 18.18 mmoles) and epichlorohydrin(14.22 mL, 181.8 mmoles) was heated to 40° C. After 2.5 h, the reactionwas cooled down to RT and the excess epichlorohydrin was evaporated invacuo. Sulfuric acid (5.52 mL) was added slowly to the crude mixture.After completion of addition, the reaction flask was placed in apreheated oil bath (150° C.). The reaction mixture was heated for 30minutes, cooled down to RT, and quenched with ice. The aqueous layer wasbasified with 10% KOH and extracted with EtOAc (3×300 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated to yield a crudeoil. This oil was purified using silica gel chromatography (70:28:2hexanes:DCM: 2M NH₃ in EtOH) to obtain the oxazepine 13 (1.47 g, 6.13mmole, 34%). LRMS: 239.

Synthesis of 4-benzyl-2-(4-trifluoromethyl-phenoxymethyl)-[1,4]oxazepane

To a solution of KOH (131 mg, 2.3 mmoles) in DMSO (2 mL) was added4-trifluoromethylphenol (189 mg, 1.17 mmoles) followed by the halide 13(280 mg, 1.17 mmoles). After completion of addition, the reactionmixture was heated to 55° C. After 12 h, the reaction mixture was cooledto RT and quenched with water. The aqueous layer was extracted withEtOAc (3×2 mL) and the combined organic layers were dried over Na₂SO₄and concentrated to yield a crude oil. The crude material was purifiedusing silica gel chromatography (9:1 hexane:EtOAc-85:15 hexanes:EtOAc)to yield 14 (113.5 mg, 0.31 mmole, 26%). LRMS: M+366.

Synthesis of 2-(4-trifluoromethyl-phenoxymethyl)-[1,4]oxazepane

Benzyl-protected amine 14 (92.5 mg, 0.253 mmole) was dissolved in MeOH(8.0 mL). To this solution 10% Pd/C (78 mg) was added. The system wasalternately evacuated and filled with hydrogen from a balloon. Thereaction mixture was stirred vigorously under hydrogen for 5 h. Thesystem was purged with nitrogen and the reaction mixture was filtered.The filtrate was concentrated to yield 15 was a yellow oil (43.6 mg,0.159 mmole, 63%). LRMS: M+275.

Synthesis of[1-(4-Chloro-phenyl)-cyclobutyl]-[2-(4-trifluoromethyl-phenoxymethyl)-[1,4]oxazepan-4-yl]-methanone

To a stirred solution of 15 (43 mg, 0.158 mmole) and1-(4-chloro-phenyl)-cyclobutane carboxylic acid (50 mg, 0.237 mmole) inanhydrous dichloromethane (4 mL) was added di-isopropyl ethyl amine(82.6 μL, 0.474 mmoles) dropwise. After completion of addition, solidPyBroP (110.5 g, 0.237 moles) was added to the stirring reactionmixture. The reaction mixture continued stirring at RT for 10 h, and wasquenched with 10% KOH (aq.). The aqueous layer was extracted with EtOAc(3×8 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated to yield an oil. This crude material was purified usingsilica gel chromatography (4:1 hexane:EtOAc-1:1 hexane EtOAc) to yield16 as a brown oil (53.3 mg, 0.113 mmole, 72%). LRMS: M+468.

Synthesis of4-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-[2-(4-trifluoromethyl-phenoxymethyl)-[1,4]oxazepane

A solution of amide 16 (53.3 mg, 0.114 mmole) in anhydrous toluene (1.5mL) was cooled to 0° C. RedAl (80.54 mg, 0.399 mmole) was added dropwiseto the cooled stirring reaction mixture. After completion of addition,the reaction continued stirring at RT. After 12 h, 10% KOH was added tothe reaction mixture. The aqueous layer was extracted with EtOAc (3×2mL). The combined organic layers were dried over Na₂SO₄ and concentratedto yield an oil. The crude material was purified using silica gelchromatography (85:15 hexanes:EtOAc) to yield 11 (30 mg, 0.066 mmole,59%) LRMS: M+453.

Example 8 Synthesis of1-[1-(4-Chlorophenyl)cyclobutyl]-2-(3-phenoxymethylpiperidin-1-yl)ethanone

A mixture of 17 (0.942 g, 4.48 mmol) and thionyl chloride (2 mL) wereheated at reflux for 3 h. The reaction mixture was concentrated, dilutedwith THF (2 mL), and concentrated in vacuo to give a brownish-yellowoil. The oil was dissolved in THF (15 mL) and then cooled to 0° C. Next,diazomethane (generated at 0° C. from 2 g1-methyl-3-nitro-1-nitrososguanidine in 15 mL diethyl ether and 1.36 gsodium hydroxide in 15 mL water) was added. The resulting solution wasmaintained at 0° C. overnight. Hydrochloric acid (5 mL; 4 M) wascarefully added. The reaction mixture was maintained at 0° C. for 1 h,and then concentrated to a yellow oil. The oil was purified by columnchromatography on silica gel eluting with hexane/ethyl acetate (90:10)to give 18 as a colorless oil.

To a solution of 18 (96 mg, 0.393 mmol) in acetone (0.5 mL) was addedsodium iodide (59 mg, 0.393 mmol). After 5 min at room temperature, themixture was added to a mixture of 19 (100 mg, 0.328 mmol) and potassiumcarbonate (226 mg) in acetone (0.5 mL). The resulting mixture was heatedat 50° C. for 18 h. The reaction mixture was poured into water (20 mL)and extracted with ethyl acetate (2×20 mL). The organic extracts werecombined, washed with brine (15 mL), dried over anhydrous sodiumsulfate, filtered, and concentrated to a yellow oil. The oil waspurified by column chromatography on silica gel eluting withhexane/ethyl acetate/2 N ammonia in ethanol (80:16:4) to give 20 as acolorless oil.

Example 9 Synthesis of1-[1-(4-Chloro-phenyl)cyclobutyl]-2-(3-phenoxymethylpiperidin-1-yl)ethanol

To a solution of 20 (56 mg, 0.141 mmol) in methanol (1 mL) at 0° C. wasadded sodium borohydride (11 mg, 0.282 mmol). The reaction mixture wasmaintained at room temperature for 2 h. The reaction mixture was pouredinto water (10 mL) and extracted with ethyl acetate (2×15 mL). Theorganic extracts were combined, washed with brine (10 mL), dried overanhydrous sodium sulfate, filtered, and concentrated to give a colorlessoil. The oil was purified by column chromatography on silica gel elutingwith hexane:ethyl acetate:2 N ammonia in ethanol (80:16:4) to give 21 asa colorless oil.

Synthesis of1-[1-(4-Chlorophenoyl)cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)piperidin-1-yl]ethanone

To a solution of 18 (96 mg, 0.396 mmol) in acetone (1.0 mL) was addedsodium iodide (59 mg, 0.393 mmol). After 5 min at room temperature, themixture was added to a mixture of 22 (123 mg, 0.330 mmol) and potassiumcarbonate (228 mg) in 0.5 mL acetone. The resulting mixture was heatedat 50° C. for 18 h. The reaction mixture was poured into water (20 mL)and extracted with ethyl acetate (2×20 mL). The organic extracts werecombined, washed with brine (15 mL), dried over anhydrous sodiumsulfate, filtered, and concentrated to give a yellow oil. The oil waspurified by column chromatography on silica gel eluting withhexane:ethyl acetate:2 N ammonia in ethanol (80:16:4) to give 23 as acolorless oil.

Example 11 Synthesis of1-[1-(4-Chlorophenyl)cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)piperidin-1-yl]ethanol

To a solution of 23 (121 mg, 0.26 mmol) in 2 mL methanol at roomtemperature was added sodium borohydride (20 mg, 0.52 mmol). Thereaction mixture was maintained at room temperature for 2 h. Thereaction mixture was poured into water (20 mL), and extracted with ethylacetate (2×20 mL). The organic extracts were combined, washed with brine(15 mL), dried over anhydrous sodium sulfate, filtered, and concentratedto a colorless oil. The oil was purified by column chromatography onsilica gel eluting with hexane:ethyl acetate:2 N ammonia in ethanol(80:16:4) to give 24 as a colorless oil.

Example 12 Synthesis of N-1-Carbobenzyloxy[3-R-(2′-anilinocarboxy]piperidine

A solution of R-Cbz-nipecotic acid (0.038 mol, 10.0 g) and aniline (4.0equiv, 0.15 mmol, 14 mL) in CH₂Cl₂ at 0° C. was treated with DCC (1.5equiv, 0.057 mol, 12.0 g) under Ar. The reaction mixture was allowed towarm to 25° C. and stirred for 12 h. The reaction mixture was thenfiltered to remove the dicyclohexyl urea, and the solvent was removed invacuo. Chromatography (SiO₂, 2.5 cm×30.5 cm, 1:1 hexane-EtOAc) provided25 (10.0 g, 12.8 g theoretical, 78%) as a white foam: R_(f) 0.45 (SiO₂,1:1 hexane-EtOAc): LRMS m/z 338 (M⁺, C₂₀H₂₂N₂O₃, requires 338).

Example 13 Synthesis of Piperidine-3-R-carboxylic Acid Phenylamide

A solution of 25 (0.015 mol, 5.0 g) and PdC 30% (100 mg) in CH₃OH at 25°C. was added to a Paar hydrogenator low pressure reaction vessel. Themixture was reacted at 55 psi with vigorous shaking until hydrogenuptake subsided (2 h). The catalyst was filtered through a pad ofCelite. The filtrate was concentrated in vacuo which provided 26 (3.0 g,3.0 g theoretical, 99%) as a white foam: LRMS m/z 204 (M⁺, C₁₂H₁₆N₂O,requires 204).

Example 14 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutanecarbonyl]-piperidine-3-R-carboxylicAcid Phenylamide

A solution of 26 (2.95 mmol, 603 mg), 1-(4-chlorophenyl)-1-cyclobutanecarboxylic acid (1.5 equiv, 4.43 mmol, 932 mg) and iPr₂NEt (3.0 equiv,8.85 mmol, 1.5 mL) in CH₂Cl₂ (10 mL) was treated with PyBroP (1.5 equiv,4.43 mmol, 2.07 g) under Ar at 0° C. After warming to 25° C., andstirring for 12 h, the reaction mixture was quenched with 10% aqueousHCl and extracted with EtOAc (3×25 mL). The organic layer was thenwashed with NaHCO_(3(sat)) and dried with NaCl_((sat)) and MgSO_(4(s)).Chromatography (SiO₂, 2.5 cm×30.5 cm, 2:1 hexane-EtOAc) provided 27(0.851 g, 1.17 g theoretical, 73%) as a white foam: R_(f) 0.17 (SiO₂,2:1 hexane-EtOAc); LRMS m/z 396 (M⁺, C₂₃H₂₅ClN₂O₂, requires 396).

Example 15 Synthesis of{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-R-ylmethyl}-phenyl-amine

A solution of 27 (0.504 mmol, 200 mg) in toluene (2 mL) at 0° C. wastreated with 3.0 M Red-Al (3.5 equiv, 1.76 mmol) under Ar. The reactionmixture stirred for 12 h, and returned to 25° C. The reaction mixturewas then cooled to 0° C., quenched with 10% aqueous NaOH and extractedwith EtOAc (3×25 mL). The organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The reaction mixture was purified by chromatography (PTLC,SiO₂, 20 cm×20 cm, 1 mm, 2:1 hexane-EtOAc) which provided 28 (170 mg,186 mg theoretical, 91%) as a colorless oil: R_(f) 0.61 (SiO₂, 2:1hexane-EtOAc); LRMS m/z 368 (M⁺, C₂₃H₂₉ClN₂, requires 368).

Example 16 Synthesis of 3-Phenoxymethyl-piperidine-1-carboxylic AcidTert-Butyl Ester

A solution of 3-hydroxymethyl-piperidine-1-carboxylic acid tert-butylester (4.64 mmol, 1.00 g), phenol (3.0 equiv, 13.92 mmol, 1.2 mL) andtriphenylphosphine (3.0 equiv, 13.92 mmol, 3.65 g) in THF at 0° C. wastreated with DEAD (3.0 equiv, 13.92 mol, 2.2 mL) under Ar. The reactionmixture was allowed to warm to 25° C., and stirred for 5 h. The reactionmixture was quenched with 10% NaOH (20 mL) and then extracted with EtOAc(2×25 mL). The combined organics were dried with NaCl_(4(sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(SiO₂, 2.5 cm×30.5 cm, 6:1 hexane-EtOAc) provided 29 (0.626 g, 1.35 gtheoretical, 46%) as a white solid: R_(f) 0.46 (SiO₂, 6:1 hexane-EtOAc);LRMS m/z 291 (M⁺, C₁₇H₂₅NO₃, requires 291).

Example 17 Synthesis of 3-Phenoxymethyl-piperidine

A solution of 29 (0.343 mmol, 100 mg) in CH₂Cl₂ (500 μL) at 0° C. wastreated with TFA (500 μL). The reaction mixture was allowed to warm to25° C., and stirred for 1 h. The solvent was removed under a stream ofN₂ which provided 30 (105 mg, 105 mg theoretical, 99%) as a white solid:LRMS m/z 192 (M⁺, C₁₂H₁₈NO⁺, requires 192).

Example 18 Synthesis of1-(4-Chloro-phenyl)-2-(3-phenoxymethyl-piperidin-1-yl)-ethanone

A solution of 3-phenoxymethyl-piperidine 30 (0.343 mmol, 105 mg),2-bromo-4′-chloroacetophenone (31) (1.5 equiv, 0.515 mmol, 120 mg) andK₂CO₃ (3.0 equiv, 1.03 mmol, 142 mg) in CH₃CN was heated to 60° C. andstirred for 12 h. The reaction mixture was quenched with H₂O (10 mL),and then extracted with EtOAc (2×15 mL). The combined organics weredried with NaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed invacuo and chromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm, 9:1hexane-acetone) provided 32 (106 mg, 118 g theoretical, 46%) as acolorless oil: R_(f) 0.52 (SiO₂, 9:1 hexane-acetone); LRMS m/z 344 (M⁺,C₂₀H₂₂ClNO₂, requires 344).

Example 19 Synthesis of1-(4-Chloro-phenyl)-2-(3-phenoxymethyl-piperidin-1-yl)-ethanol

A solution of 32 (0.259 mmol, 89 mg) in CH₃OH was treated with NaBH₄(3.0 equiv, 0.777 mmol, 30 mg) at 0° C. and stirred for 2 h. Thereaction mixture was quenched with 10% HCl (5 mL) and then neutralizedwith NaHlCO_(3(sat)) and extracted with EtOAc (2×10 mL). The combinedorganics were dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm,9:1 EtOAcCH₃OH) provided 33 (72 mg, 90 mg theoretical, 80%) as acolorless oil: R_(f) 0.60 (SiO₂, 9:1 EtOAc—CH₃OH); LRMS m/z 346 (M⁺,C₂₀H₂₄ClNO₂, requires 346).

Example 20 Synthesis of3-Phenoxymethyl-1-(1-phenyl-cyclobutylmethyl)-piperidine

A solution of 30 (0.343 mmol, 105 mg) and1-phenyl-cyclobutanecarbaldehyde (1.5 equiv, 0.515 mmol, 83 mg) inbenzene were heated to reflux utilizing a Dean-Stark trap. The benzenewas replaced three times. After the last reflux period, the solventswere removed in vacuo. The resulting oil was dissolved in THF (1 mL) andtreated with NaCNBH₃ (3.0 equiv, 1.03 mmol, 65 mg) andtrimethylorthoformate (1 mL) at 25° C. After stirring for 12 h, thereaction mixture was quenched with 10% HCl (5 mL) and then neutralizedwith NaHCO_(3(sat)) and extracted with EtOAc (2×10 mL). The combinedorganics were dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo, and chromatography (PTLC, SiO₂, 20 cm×20 cm, 1mm, 3:1 hexanes-EtOAc) provided 34 (68 mg, 115 mg theoretical, 59%) as acolorless oil: R_(f) 0.54 (SiO₂, 3:1 hexanes-EtOAc); LRMS m/z 335 (M⁺,C₂₃H₂₉NO, requires 335).

Example 21{1-[1-(4-Chloro-phenyl-cyclobutylmethyl]-piperidin-3-R-ylmethyl}-methyl-phenyl-amine

A solution of the 28 (0.228 mmol, 84 mg) in THF (1 mL) at −78° C. wastreated with 1.6 M nBuLi (1.5 equiv, 0.342 mmol, 214 μL) under Ar. Thereaction mixture was warmed to 0° C. for 30 min and then cooled again to−78° C. CH₃I (1.5 equiv, 0.342 mmol, 21 μL) was then added and thereaction mixture stirred at 0° C. for 5 min. The reaction was quenchedwith NaHCO_(3(sat)) and extracted with EtOAc. The combined organics weredried with NaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed invacuo and chromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm, 3:1hexanes-EtOAc) provided 35 (87 mg, 87 mg theoretical, 99%) as a yellowoil: R_(f) 0.38 (SiO₂, 3:1 hexanes-EtOAc); LRMS m/z 383 (M⁺, C₂₄H₃₁ClN₂,requires 383).

Example 22 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-[2-(4-trifluoromethyl-phenoxy)-ethyl]-piperidine

To a solution of N-Boc-3-piperidine acetic acid (500 mg, 2.1 mmol) inTHF (10 mL) at room temperature was added BH₃-THF complex (5.1 mL of a1.0 M solution, 5.1 mmol) dropwise. The reaction was allowed to stir atthis temperature for two-and-a-half hours before quenching by theaddition of 2 M HCl (approx. 10 mL). This mixture was allowed to stirfor fifteen minutes before neutralizing by the addition of 2 M NaOH andextracting with ethyl acetate. The organic layer was dried (MgSO₄),filtered and concentrated in vacuo to provide the desired product 36(488 mg, 100%) which required no further purification. LRMS calculatedfor C₁₂H₂₃NO₃ 229.17, found 229.71.

To a solution of primary alcohol 36 (300 mg, 1.3 mmol) indichloromethane (6 mL) at room temperature was added iPr₂NEt (0.57 mL,3.3 mmol) followed by MsCl (0.11 mL, 1.4 mmol). The reaction mixture wasallowed to stir for one hour before concentrating and purifying theresulting residue by flash column chromatography using a gradient of 30to 50% ethyl acetate/petroleum ether to provide the desired mesylate(not shown) (336 mg, 83%). LRMS calculated for C₁₃H₂₅NO₅S 307.15, found307.34. To the mesylate (336 mg, 1.1 mmol) in DMF (5 mL) was addedα,α,α-trifluoro-p-cresol (355 mg, 2.2 mmol) followed by Cs₂CO₃ (1.8 g,5.5 mmol). The reaction mixture was heated to 75° C. for thirty minutesbefore cooling to room temperature, diluting with ethyl acetate andwashing several times with brine. The organic layer was then dried(MgSO₄), filtered and concentrated in vacuo. The resulting residue waspurified by flash column chromatography using a gradient of 6 to 10%acetone/hexane to provide 37 (342 mg, 84%). LRMS calculated forC₁₉H₂₆F₃NO₃ 373.19, found 373.96.

N-Boc-protected 37 (342 mg, 0.92 mmol) was then stirred in 40% TFA/DCM(5 mL) for one hour before concentrating in vacuo. The resulting residuewas diluted with dichloromethane, washed with saturated aqueous sodiumbicarbonate. The organic layer was then dried (MgSO₄), filtered andconcentrated in vacuo. To the resulting free amine (not shown) indichloromethane (5 mL) was then added 1-(4-chlorophenyl)-1-cyclobutanecarboxylic acid (289 mg, 1.4 mmol) and iPr₂NEt (0.80 mL, 4.6 mmol)followed by PyBroP (641 mg, 1.4 mmol). The resulting solution wasallowed to stir overnight at room temperature before quenching with 10%KOH and washing with ethyl acetate. The organic layer was then dried(MgSO₄), filtered, concentrated in vacuo and the resulting residuepurified by flash column chromatography using a gradient of 20 to 30%ethyl acetate/hexane to provide amide 38 (238 mg, 56% for two steps).LRMS calculated for C₂₅H₂₇ClF₃NO₂ 465.17, found 466.18.

To amide 38 (100 mg, 0.22 mmol) in toluene (1 mL) was cautiously addedRed-Al (0.23 mL, 0.75 mmol). The resulting solution was allowed to stirat room temperature for one hour before diluting with ethyl acetate andquenching with 10% aqueous KOH. The layers were separated and theaqueous layer further washed with ethyl acetate. The combined organiclayers were then dried (MgSO₄), filtered, concentrated in vacuo and theresulting residue purified by flash column chromatography using 1% 2MNH₃ in EtOH/DCM to provide amine 39 (54 mg, 56%). LRMS calculated forC₂₅H₂₉ClF₃NO 451.19, found 451.48. ¹H NMR (300 MHz, CDCl₃): δ 7.53 (d,J=8.5 Hz, 2H), 7.18 (d, J=8.4 Hz, 2H), 7.07 (d, J=8.3 Hz, 2H), 6.89 (d,J=8.54 Hz, 2H), 3.72-3.83 (m, 2H), 2.40-2.65 (m, 2H), 2.38 (m, 1H),1.95-2.26 (m, 71, 1.39-1.88 (m, 8H), 0.80-0.93 (m, 1H). ¹³C NMR (75 MHz,CDCl₃): δ 161.4, 148.5, 137.4, 130.8, 127.7, 127.6, 126.8, 122.8 (m),114.4, 68.6, 66.0, 61.7, 56.4, 46.9, 33.2, 33.1, 31.6, 31.4, 30.5, 25.2,15.9.

Example 23 Synthesis of1-[2-(chloro-phenyl)-2-methyl-propyl]-3-(4-trifluoromethyl-phenoxymethyl)-piperidine

To 3-piperidine methanol (1.0 g, 8.7 mmol) in dichloromethane (40 mL)was added 2-(4-chlorophenyl)-2-methyl propionic acid (2.6 g, 13.0 mmol)and iPr₂NEt (4.5 mL, 26.6 mmol) followed by PyBroP (6.1 g, 13.0 mmol).The resulting solution was allowed to stir overnight at room temperaturebefore diluting with ethyl acetate and quenching with 10% KOH. Thelayers were separated and the aqueous layer further washed with ethylacetate. The combined organic layers were then dried (MgSO₄), filteredand concentrated in vacuo. The resulting residue was purified by flashcolumn chromatography using a gradient of 40 to 50% ethylacetate/petroleum ether to provide amide 40 (1.92 g, 75%). LRMScalculated for C₁₆H₂₂ClNO₂ 295.13, found 295.85.

To a solution of 40 (536 mg, 1.8 mmol) in dichloromethane (8 mL) at roomtemperature was added iPr₂NEt (0.79 mL, 4.5 mmol) followed by MsCl (0.15mL, 2.0 mmol). The reaction mixture was allowed to stir for one hourbefore concentrating and purifying the resulting residue by flash columnchromatography using a gradient of 30 to 50% ethyl acetate/petroleumether to provide the desired mesylate (not shown) (569 mg, 84%). LRMScalculated for C₁₇H₂₄ClNO₄S 373.11, found 374.45. To the mesylate (569mg, 1.5 mmol) in DMF (7 mL) was added α,α,α-trifluoro-p-cresol (259 mg,1.6 mmol) followed by Cs₂CO₃ (1.5 g, 4.6 mmol). The reaction mixture washeated to 75° C. for two hours before cooling to room temperature,diluting with ethyl acetate and washing several times with brine. Theorganic layer was then dried (MgSO₄), filtered and concentrated invacuo. The resulting residue was purified by flash column chromatographyusing 30% ethyl acetate/hexane to provide ether 41 (477 mg, 71%). LRMScalculated for C₂₃H₂₅ClF₃NO₂ 439.15, found 440.27.

To 41 (100 mg, 0.23 mmol) in toluene (1 mL) was cautiously added Red-Al(0.24 mL, 0.80 mmol). The resulting solution was allowed to stir at roomtemperature for one hour before adding an additional portion of Red-Al(0.10 mL, 0.34 mmol) and stirring at room temperature overnight. Thereaction was then diluted with ethyl acetate and quenched with 10%aqueous KOH. The layers were separated and the aqueous layer furtherwashed with ethyl acetate. The combined organic layers were then dried(MgSO₄), filtered, concentrated in vacuo and the resulting residuepurified by flash column chromatography using 0.5% 2M NH₃ in EtOH/DCM toprovide amine 42 (52 mg, 54%). LRMS calculated for C₂₃H₂₇ClF₃NO 425.17,found 425.78. ¹H NMR (300 MHz, CDCl₃): 7.57 (d, J=8.7 Hz, 2H), 7.35 (d,J=8.8 Hz, 2H), 7.27 (d, J=8.7 Hz, 2H), 6.93 (d, J=8.5 Hz, 2H), 3.72-3.84(m, 2H), 2.53-2.55 (m, 1H), 2.41 (m, 3H), 2.13-2.21 (m, 1H), 2.01-2.03(m, 2H), 1.48-1.71 (m, 3H), 1.32 (s, 6H), 1.06-1.19 (m, 1H).

Example 24 Synthesis of[3-(Benzo[1,3]-dioxol-5-yloxymethyl)-piperidin-1-yl]-[1-(4-chloro-phenyl)-cyclobutyl]-methanone(44)

Methanesulfonic Acid1-[1-(4-chloro-phenyl)-cyclobutanecarbonyl]-piperidin-3-ylmethyl Ester(43)

To a solution of 3-piperidinemethanol (5.0 g, 43.4 mmol),1-(4-chlorophenyl)-1-cyclobutanecarboxylic acid (9.14 g, 43.4 mmole),and diisopropylethylamine (11.22 g, 86.8 mmol) in dichloromethane (100mL) at 0° C. was added PyBroP® (22.26 g, 47.8 mmol). The reaction wasstirred at 0° C. for 1 h and then at room temperature for 4 h. Thereaction mixture was washed successively with water, 1 N HCl, water,sat. sodium bicarbonate solution, and water (100 m-L each). The organiclayer was dried over anhydrous sodium sulfate, filtered and concentratedby rotary evaporation. The residue was purified by chromatography onsilica gel, eluting with dichloromethane/methanol (96:4) to give 5.8 gof the amide 1 as a thick gum.

The amide 1 (5.0 g) was dissolved in dichloromethane (50 mL) and cooledto 0° C. To this solution was added pyridine (5.0 mL) followed bydropwise addition of methanesulfonyl chloride (2.05 g, 17.9 mmol). Thereaction was stirred at for 1 h and then at room temperature overnight.The reaction mixture was washed successively with water, 1 N HCl, water,sat. sodium bicarbonate solution, and water (100 mL each). The reactionmixture was washed successively with water, sat. sodium bicarbonatesolution, and water (100 mL each). The organic layer was dried overanhydrous sodium sulfate, filtered and concentrated by rotaryevaporation. The residue was purified by chromatography on silica gel,eluting with hexane/ethyl acetate (2:1) to give 4.45 g of 43 as a tangum. C₁₈H₂₄ClNO₄S, MS (m/z)=386 (MH+).

[3-(Benzo[1,3-dioxol-5-yloxymethyl)-piperidin-1-yl]-[1-(4-chloro-phenyl)-cyclobutyl]-methanone(44)

To a solution of 43 (1.0 g, 2.59 mmol) in acetonitrile (25 mL) was addedsesamol (0.36 g, 2.59 mmol) and cesium carbonate (1.27 g, 3.89 mmol).The reaction was stirred and refluxed for 20 h. After cooling to roomtemperature, the reaction mixture was filtered and most of the solventwas removed by rotary evaporation. The residue was partitioned betweendichloromethane and water, and the organic layer was washed with sat.sodium carbonate (2×50 mL) and water. The organic layer was dried overanhydrous sodium sulfate, filtered and concentrated by rotaryevaporation. The residue was purified by chromatography on silica gel,eluting with dichloromethane/methanol (98:2) to give 0.68 g of the amide44 as a crystalline solid. C₂₄H₂₆ClNO₄, MS (m/z)=428 (MH+).

Example 25 Synthesis of 1-Phenylcyclobutylcarboxaldehyde (47)

A solution of 3.018 g (19.2 mmol) of 1-phenylcyclobutanecarbonitrile in60 mL of toluene was cooled to −70° C. and 38 mL of 1 M DIBAL-H inhexane was added dropwise in 30 min. The mixture was stirred at −70° C.for 30 min and at ambient temperature for 4 hours, whereupon 3 mL ofethyl formate was added and stirring was continued for 1 hour. Themixture was poured into saturated ammonium chloride solution (70 mL);after 30 min, 2M aqueous sulfuric acid (100 mL) was added and theproduct was isolated with ether (3×75 mL). The organic phase was driedover MgSO₄ and filtered. Evaporation to dryness furnished the crudeproduct which was purified by column chromatography leading to 2.5 galdehyde 47, 83% yield. ¹H NMR (CDCl₃) 1.92-2.13 (m, 2H), 2.40-2.51 (m,2H), 2.74-2.83 (m, 2H), 7.18-7.22 (m, 2H), 7.28-7.34 (m, 1H), 7.39-7.45(m, 2H), 9.58 (s, 1H). ¹³C NMR (CDCl₃) 16.1, 28.6, 57.8, 126.7, 127.3,129.1, 141.2, 199.7.

Example 26 Synthesis of 1-(4-Methoxyphenyl)cyclobutanecarbonitrile (48)

A solution of 3.27 g (22.2 mmol) of (4-methoxyphenyl)acetonitrile and4.93 g (24.4 mmol) of 1,3-dibromopropane in 15 mL of ether was addeddropwise into 1.17 g (48.8 mmol) of NaH in 60 mL of DMSO. Thetemperature was held between 250 and 35° by water bath cooling. Themixture was stirred at room temperature for 18 hours. The mixture wascooled in ice water and 3 mL of 2-propanol was added dropwise, followedby the addition of 50 mL of water. The mixture was extracted with hexane(3×100 mL), and the combined extracts were washed with water (3×75 mL),dried over anhydrous magnesium sulfate, filtered, and concentrated toyield 2.6 g (62%) product as colorless oil. ¹H NMR (CDCl₃) δ 2.01-2.14(m, 1H), 2.35-2.50 (m, 1H), 2.55-2.66 (m, 2H), 2.77-2.87 (m, 2H), 3.83(s, 3H), 6.94 (d, J=9.0 Hz, 2H), 7.36 (d, J=9.0 Hz, 2H). ¹³C NMR (CDCl₃)δ 17.3, 35.1, 39.9, 55.6, 114.5, 124.9, 127.0, 132.1, 159.4.

Synthesis of 1-(4-Chlorophenyl)cyclobutanecarbonitlile (49)

A solution of 3.37 g (22.2 mmol) of 4-Chlorobenzyl cyanide and 4.93 g(24.4 mmol) of 1,3-dibromopropane in 15 mL of ether was added dropwiseinto 1.17 g (48.8 mmol) of NaH in 60 mL of DMSO. The temperature washeld between 25° and 350 by water bath cooling. The mixture was stirredat room temperature for 18 hours. The mixture was cooled in ice waterand 3 mL of 2-propanol was added dropwise, followed by the addition of50 mL of water. The mixture was extracted with hexane (3×100 mL), andthe combined extracts were washed with water (3×75 mL), dried overanhydrous magnesium sulfate, filtered, and concentrated to yield 2.9 g(70%) product as colorless oil. ¹H NMR (CDCl₃) δ 2.04-2.15 (m, 1H),2.39-2.52 (m, 1H), 2.55-2.65 (m, 2H), 2.79-2.88 (m, 2H), 7.38 (s, 4H).¹³C NMR (CDCl₃) 17.3, 34.9, 40.0, 124.2, 127.3, 129.4, 134.1, 138.6.

Synthesis of 1-(3-Chlorophenyl)cyclobutanecarbonitrile (50)

A solution of 3.37 g (22.2 mmol) of 3-Chlorobenzyl cyanide and 4.93 g(24.4 mmol) of 1,3-dibromopropane in 15 mL of ether was added dropwiseinto 1.17 g (48.8 mmol) of NaH in 60 mL of DMSO. The temperature washeld between 25° and 350 by water bath cooling. The mixture was stirredat room temperature for 18 hours. The mixture was cooled in ice waterand 3 mL of 2-propanol was added dropwise, followed by the addition of50 mL of water. The mixture was extracted with hexane (3×100 mL), andthe combined extracts were washed with water (3×75 mL), dried overanhydrous magnesium sulfate, filtered, and concentrated to yield 3.0 g(71%) product as colorless oil. ¹H NMR (CDCl₃) δ 2.04-2.16 (m, 1H),2.38-2.50 (m, 1H), 2.56-2.67 (m, 21), 2.79-2.89 (m, 2H), 7.29-7.36 (m,3H), 7.41-7.43 (m, 1H). ¹³C NMR (CDCl₃) 17.3, 34.8, 40.1, 124.2, 126.2,128.4, 130.5, 135.2, 142.0.

Synthesis of 1-(4-Fluorophenyl)cyclobutanecarbonitrile (65)

A solution of 3.00 g (22.2 mmol) of 4-fluorophenylacetonitrile and 4.93g (24.4 mmol) of 1,3-dibromopropane in 15 mL of ether was added dropwiseinto 1.17 g (48.8 mmol) of NaH in 60 mL of DMSO. The temperature washeld between 250 and 35° by water bath cooling. The mixture was stirredat room temperature for 18 hours. The mixture was cooled in ice waterand 3 mL of 2-propanol was added dropwise, followed by the addition of50 mL of water. The mixture was extracted with hexane (3×100 mL), andthe combined extracts were washed with water (3×75 mL), dried overanhydrous magnesium sulfate, filtered, and concentrated to yield 2.35 g(61%) product as colorless oil. ¹H NMR (CDCl₃) δ 2.03-2.16 (m, 1H),2.38-2.53 (m, 1H), 2.58-2.67 (m, 21), 2.80-2.90 (m, 2H), 7.11 (dd,J=9.0, 8.4 Hz, 2H), 7.41 (dd, J=9.1, 5.1 Hz, 2H). ¹³C NMR (CDCl₃) 17.3,35.0, 39.9, 116.1 (d, J=21.5 Hz), 124.5, 127.7, 135.9, 162.5 (d, J=230Hz).

Example 27 Synthesis of1-[1-(4-Chloro-phenyl)cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-azepane

1-Benzyl-azepan-2-one

To a stirring 0° C. suspension of NaH (18.3 g, 763 mmol) in THF (195 mL)was added by addition funnel azepan-2-one (75.0 g, 667 mmol) in THF. Anadditional 2 L of solvent was added as the reaction progressed in orderto maintain agitation of the very viscous reaction suspension. Followingaddition, the reaction was allowed to warm to room temperature, and whenthe evolution of H₂ gas ceased after stirring overnight, benzyl bromidewas added dropwise by addition funnel and the reaction was stirredovernight. The crude product was filtered through Celite andconcentrated in vacuo. Recrystallization from hexanes and ethyl acetateprovided pure 1-benzyl-azepan-2-one as a white fluffy solid. ¹H NMR(CDCl₃, 300 MHz) 7.38-7.24 (5H, m), 4.61 (2H, s), 3.33-3.29 (2H, m),2.65-2.61 (2H, m), 1.77-1.66 (4H, m), 1.56-1.46 (2H, m) ppm. ¹³C NMR(CDCl₃, 75 MHz) 176.13, 138.06, 128.67, 128.31, 127.43, 51.20, 49.04,37.33, 30.12, 28.26, 23.58 ppm.

1-Benzyl-2-oxo-azepane-3-carboxylic Acid Methyl Ester

LDA was prepared as follows: Diisopropyl amine (15.2 mL, 110 mmol)freshly distilled under N₂ and over CaH₂ was added to 110 mL ofanhydrous THF in a dry flask, and the solution was cooled to 0° C. in anice bath. nBuLi (73.3 mL, 111 mmol) was added dropwise, and the reactionwas stirred at 0° C. for 1 hour. The freshly prepared LDA was addeddropwise to a −70° C. solution of 1-benzyl-azepan-2-one (10.98 g, 544.0mmol) dissolved in anhydrous Et₂O (70 mL). The reaction was stirred at−70° C. for 1 hour, then dimethyl carbonate (4.55 mL, 544 mmol) wasadded dropwise. The reaction was allowed to warm to room temperatureovernight. The reaction was judged complete by HPLC, and was slowlypoured into 5N HCl stirring in an ice bath. The organic layer wasextracted. The aqueous layer was washed with CH₂Cl₂ two times, and thecombined organics were dried with Na₂SO₄ and concentrated in vacuo.Crude material was purified on an automated flash column with 80:20Hexanes:EtOAc to obtain 10.85 g, (77%) of1-benzyl-2-oxo-azepane-3-carboxylic acid methyl ester as a pale yellowoil. ¹H NMR (CDCl₃, 300 MHz) 7.42-7.10 (5H, broad s), 4.61 (1H, d,J=14.7 Hz), 4.50 (1H, d, J=14.7 Hz), 3.74 (3H, s), 3.7-3.64 (1H, m),3.40-3.13 (2H, m), 2.12-1.98 (1H, m), 1.92-1.73 (2H, m), 1.66-1.42 (2H,m), 1.32-1.16 (1H, m) ppm. ¹³C NMR (CDCl₃, 75 MHz) 171.94, 171.04,137.28, 128.55, 128.24, 127.43, 52.15 (2), 51.27, 48.31, 27.87, 27.41,25.91 ppm. LRMS: 261.73.

(1-Benzyl-azepan-3-yl)-methanol

1-Benzyl-2-oxo-azepan-2-carboxylic acid methyl ester (0.2154 g, 0.8243mmol) dissolved in anhydrous THF (2.9 mL) was added to a stirringsuspension of LiAlH₄ in THF (1.5 mL) over approx. 1.5 hours. Thereaction was stirred overnight. The reaction was judged complete by TLCand was quenched by the sequential addition of H₂O (0.4 mL), then 2NNaOH (1.0 mL) and H₂O (0.4 mL). The reaction was stirred at roomtemperature for 30 minutes, then was filtered, dried with Na₂SO₄, andconcentrated in vacuo. Crude material was purified by automated silicagel chromatography with 15:85:5 CH₂Cl₂: Hexanes: 2N NH₃ in ethyl alcoholto obtain 0.1062 g (59%) of pure (1-benzyl-azepan-3-yl)-methanol. ¹H NMR(CDCl₃, 300 MHz) 7.40-7.23 (5H, m), 3.65 (2H, s), 3.54 (1H, dd, J=10.4,3.5 Hz), 3.43 (1H, dd, J=10.4, 5.4 Hz), 2.82 (1H, J=13.3, 3.1 Hz), 2.77(2H, m), 2.44 (1H, ddd, J=12.2, 8.6, 3.3 Hz), 1.90-1.45 (6H, m) ppm. ¹³CNMR (CDCl₃, 75 MHz) 139.14, 128.97, 128.14, 126.91, 67.20, 63.85, 58.49,56.87, 39.59, 29.68, 29.43, 25.23 ppm. LRMS: 219.64.

Azepan-3-yl-methanol

(1-Benzyl-azepan-3-yl)-methanol (0.0922 g, 0.4192 mmol) dissolved inMeOH (1 mL) was added to a stirring suspension of 10% Pd/C (14.4 mg) in5 mL MeOH. The reaction was purged with H₂, and the reaction was stirredat room temperature overnight. The reaction was judged complete by¹H-NMR analysis of an aliquot from the reaction. The reaction wasfiltered through a pad of Celite wet with MeOH and was rinsed with MeOH,and concentrated in vacuo to obtain pure azepan-3-yl-methanol in 60%yield (0.0323 g), which was used in the next step without furtherpurification. ¹H NMR (CDCl₃, 300 MHz, partial) 3.14-2.70 (4H, m),1.92-1.73 (4H, m), 1.68-1.42 (3H, m) ppm. ¹³C NMR (CDCl₃, 75 MHz) δ7.32,52.01, 50.33, 41.31, 31.05, 29.76, 25.44 ppm.

[1-(4-Chloro-phenyl)-cyclobutyl]-(3-hydroxymethyl-azepan-1-yl)-methanone

iPrEtN (1.78 mL, 10.2 mmol) and pyBrOP (2.39 g, 5.12 mmol) were added atroom temperature to a stirring solution of azepan-3-yl-methanol (0.4392g, 3.41 mmol) and 1-(4 Chlorophenyl)-cyclobutanecarboxylic acid (1.0778g, 5.12 mmol) in CH₂Cl₂ under N₂. When the reaction was complete, thereaction was subjected to aqueous work-up. Silica gel purification (4:1to 2:3 Hexanes:Ethyl acetate) provided the desired product in 95% yield(1.0395 g). LRMS: 321.91.

[1-(4-Chloro-phenyl)-cyclobutyl]-[3-(4-trifluoromethyl-phenoxymethyl)-azepan-1-yl]-methanone

To a room temperature solution of[1-(4-Chloro-phenyl)-cyclobutyl]-(3-hydroxymethyl-azepan-1-yl)-methanone(0.3307 g, 1.15 mmol) in tetrahydrofuran (5.75 mL, 0.2M) under N₂ wasadded triphenyl phosphine (0.9050 g, 3.45 mmol) and4-trifluoromethylphenol (0.56 mL, 3.45 mmol). The solution was cooled to0° C., then diethyl azodicarboxylate (0.54 mL, 3.1 mmol) was addeddropwise over 10 minutes. When the reaction was complete by HPLC (2hours), ethyl acetate and 10% aqueous NaOH was added. The organics wereremoved, dried with Na₂SO₄, concentrated, then taken up in Hexanes/ethylacetate (70:30) and filtered to remove triphenyl phosphine oxide. Theremaining yellow oil was purified by silica gel chromatography (3:1 to2:3 Hexanes:ethyl acetate). The product was further purified by silicagel chromatography with 9:1 hexanes:ethyl acetate to obtain pure productin 26% yield (0.1301 g). Partial ¹H NMR (CDCl₃, 300 MHz) 7.55 (2H, t,J=8.7 Hz), 7.36-7.26 (4H, m), 6.92 (2H, dd, J=34.9, 8.7 Hz), 4.00-3.85(2H, m), ppm. Partial ¹³C NMR (CDCl₃, 75 MHz) 174.95, 161.66, 142.17,132.35, 129.10, 126.95, 126.77, 114.61, 114.61, 71.50, 71.10 ppm. LRMS:465.26.

1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-azepane(168)

To a stirring 0° C. solution of[1-(4-Chloro-phenyl)-cyclobutyl]-[3-(4-trifluoromethyl-phenoxymethyl)-azepan-1-yl]-methanone(0.13 g, 0.28 mmol) under N₂ in toluene (2.8 mL, 0.1M) was added sodiumbis(2-methoxyethoxy)aluminum hydride (65+weight % in toluene) (0.30 mL,0.98 mmol) dropwise with stirring. When the reaction was complete byHPLC, the reaction was quenched with H₂O. 10% NaOH and ethyl acetatewere added, and the organic was removed, dried with Na₂SO₄, andconcentrated. The crude reaction mixture was purified by silica gelchromatography (90:8:2 Hexanes:methylene chloride: 2N NH₃ in ethylalcohol). A 40% yield (0.0504 g) of pure material was obtained as wellas other impure fractions. ¹H NMR (CDCl₃, 300 MHz) 7.54 (2H, d, J=8.9Hz), 7.22 (2H, d, J=8.4 Hz), 7.09 (2H, d, J=8.3 Hz), 6.87 (2H, d, J=8.8Hz), 3.62-3.45 (2H, m), 2.89 (1H, d, J=13.7 Hz), 2.80 (1H, d, J=13.7Hz), 2.60 (1H, td, J=13.3, 3.7 Hz), 2.55-2.36 (3H, m), 2.30-2.10 (4H,m), 2.08-1.90 (2H, m), 1.90-1.25 (7H, m) ppm. ¹³C NMR (CDCl₃, 75 MHz)161.75, 148.53, 130.96, 128.00, 127.91, 127.03, 126.98, 122.98, 114.61,71.38, 70.53, 60.37, 58.87, 48.07, 39.52, 31.84, 31.64, 30.23, 29.50,24.97, 16.24 ppm. LRMS: 451.53.

Example 28 Synthesis of{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-azepan-3-yl}-methanol

To a stirring 0° C. solution of[1-(4-Chloro-phenyl)-cyclobutyl]-(3-hydroxymethyl-azepan-1-yl)-methanone(0.075 g, 0.26 mmol) under N₂ in toluene (2.6 mL, 0.1M) was added sodiumbis(2-methoxyethoxy)aluminum hydride (65+weight % in toluene) (0.28 mL,0.91 mmol) dropwise with stirring. When the reaction was complete byHPLC, the reaction was quenched with H₂O. 10% NaOH and ethyl acetatewere added, and the organic was removed, dried with Na₂SO₄, andconcentrated. The crude reaction mixture was purified by silicagel:chromatography (90:8:2 Hexanes:methylene chloride: 2N NH₃ in ethylalcohol). A 39% yield (0.0290 g) of pure material was obtained as wellas other impure fractions. ¹H NMR (CDCl₃, 300 MHz) 7.28-7.25 (2H, m),7.15-7.09 (2H, m), 3.44 (1H, dd, J=10.5, 4.5 Hz), 3.27 (1H, dd, J=10.3,5.5 Hz), 2.89 (1H, d, J=13.9 Hz), 2.83 (1H, d, J=13.9 Hz), 2.72-2.60(1H, m), 2.50-1.10 (16H, m) ppm. ¹³C NMR (CDCl₃, 75 MHz) 146.5, 131.22,128.10, 127.93, 70.95, 67.49, 61.85, 59.10, 47.62, 40.59, 32.12, 31.90,29.92, 25.41, 16.20 ppm. LRMS: 308.24.

Example 29 Synthesis of3-(4-Methoxy-phenoxymethyl)-1-(1-phenyl-cyclobutylmethyl)piperidine

To a solution of 54 (TFA salt 35.7 mg 0.11 mmol) in trimethylorthoformate (0.5 mL) was added 1-phenylcyclobutanecarboxaldehyde (18mg, 011 mmol). After stirring at room temperature for one hour, 0.1 g of(polystyrylmethyl)trimethylammonium cyanoborohydride (2.85 mmol/g) wasadded, and the reaction mixture was agitated at room temperature for 18hours. Another 18 mg of 1-phenylcyclobutanecarboxaldehyde was added intothe reaction mixture. After shaking at room temperature for 18 hours,the reaction mixture was filtered and the resin was washed with MeOH(3×0.5 mL). After conditioning a SPE column (SCX cation exchange, 0.5 gof sorbent, 2.0 mequiv/g) with MeOH (5 mL), the reaction contents wereloaded onto the column. The column was washed with MeOH (2×5 mL), andeluted with 4 mL of 2 M ammonia in MeOH. The effluent was collected intoa receiving tube, concentrated and dried in vacuo to afford 25 mg of 55,65% yield, LRMS m/z 366.

Example 30 In Vivo Evaluation of 4

Compound 4 was administered i.v. to a group of 3 ICR derived male orfemale mice (˜22 gms) and observed for the presence of acute toxicsymptoms (mortality, convulsions, tremors, muscle relaxation, sedation,etc.) and autonomic effects (diarrhea, salvation, lacrimation,vasodilation, piloerection, etc) during first 5 min (i.v). The number ofanimal deaths was observed at the subsequent 3, 24, 48, 72 hours aftercompound treatment.

Compound 4 was administered at doses of 5, 10, 20, and 30 ink/kg. Noobvious change was observed in autonomic signs of behavior for all fourdoses. After monitoring daily for 3 days, no mortality was observed.

Example 31 Antagonism of Dopamine Receptors or Transporters & FunctionalActivity

The ability of compounds of the invention to displace norephinephrineligands in vitro was determined by the methods of Galli et al. (J. Exp.Biol. 198:2197, 1995) using desipramine (IC₅₀=920 nM) as a referencecompound. The displacement of dopamine, and serotonin ligands in vitrowas determined by the methods of Gu et al. (J. Biol. Chem. 269; 7124,1994) using GBR-12909 (IC₅₀(DA uptake)=490 nM, IC₅₀ (5-HT uptake)=110nM) as a reference compound. Functional activity of the compounds wasdetermined in vitro in cellular assays using recombinant human celllines. Measurements of functional activity for serotonin uptakeinhibition was determined in human HEK-293 cell lines according to theprocedures of Gu H. et al. (J. Biol. Chem. 269: 27124, 1994) usingfluoxetine (EC₅₀=57 nM) as the reference compound. Determination offunctional activity for norephinephrine uptake inhibition wasaccomplished using a MDCK cell lines according to the methods of GalliA. et al. (J. Exp. Biol. 198:2197, 1995) with desiprmaine (EC₅₀=7 nM) asa reference compound. For determination of dopamine fictional activity,a hDAT cell line was used as described by Giros B. et al. Mol.Pharmacol. 42:383, 1992) with nomifensine (EC₅₀=11 nM) as the referencecompound.

Uptake Profile Functional Assays (IC₅₀, nM) (antagonism, EC₅₀, nM) NE DA5-HT NE DA 5-HT Compound Uptake Uptake Uptake Uptake Uptake Uptake35 >1,000 >1,000 >1,000 NA NA NA 28 <1,000 <1,000 <1,000 <1,000 <100<1,000 20 <1,000 <1,000 <1,000 NA NA NA 21 <100 <100 <1,000 NA NA NA7 >1,000 <100 >1,000 NA <100 NA 15 >1,000 >1,000 >1,000 NA NA NA16 >1,000 >1,000 >1,000 NA NA NA 11 >1,000 <1,000 >1,000 NA NA NA1 >1,000 >1,000 >1,000 NA NA NA 2 >1,000 <1,000 >1,000 NA <100 NA4 >1,000 <1,000 >1,000 NA <1,000 NA 5 NA <1,000 NA NA NA NA 33 <1,000<100 <1,000 NA NA NA 34 <1,000 <1,000 <1,000 NA NA NA 6 NA <1,000 NA NANA NA 32 <1,000 >1,000 >1,000 NA NA NA 23 <1,000 <100 >1,000 NA NA NA 24<10 <10 <1,000 NA NA NA 39 <100 <10 >1,000 NA NA NA 55 >1,000<1,000 >1,000 NA NA NA 72 NA <1,000 NA NA NA NA 76 <1,000 <1,000 >1,000NA NA NA 89 <1,000 <1000 >1,000 NA NA NA 42 <1,000 <100 >1,000 NA NA NA80 >1,000 >1,000 >1,000 NA NA NA 81 >1,000 >1,000 >1,000 NA NA NA 82<1,000 <100 <1,000 NA NA NA 8 >1,000 >1,000 NA NA NA NA 78 <1,000<1,000 >1,000 NA NA NA 79 <1,000 <1,000 <100 NA NA NA 83 <1,000 <100<1,000 <1,000 <10 <100 86 <1,000 <100 <1,000 <1,000 <10 <1,00093 >1,000 >1,000 <1,000 NA NA NA 95 >1,000 <1,000 <1,000 NA NA NA 99<1,000 <100 <1,000 <1,000 <10 >1,000 102 >1,000 <100 >1,000 <10<10 >1,000 105 <100 <10 <1,000 NA NA NA 107 <1,000 <1,000 >1,000 NA NANA 110 <1,000 <100 <1,000 <1,000 <10 >1,000 113 <1,000 <100 <1,000<1,000 <10 >1,000 114 <1,000 <100 >1,000 <1,000 <10 >1,000 115 <1,000<100 >1,000 <1,000 <10 >1,000 118 <1,000 <10 >1,000 <100 <10 >1,000 119<100 <10 >1,000 <100 <10 >1,000 122 >1,000 <1,000 >1,000 NA NA NA 128<1,000 <100 <1,000 <1,000 <10 <1,000 123 <1,000 <10 <1,000 <1,000 <10<1,000 129 <100 <10 >1,000 <100 <10 <100 130 <1,000 <100 <1,000 <100<10 >1,000 82 <1,000 <100 <1,000 <1,000 <100 NA 131 <1,000 <100 <1,000NA NA NA 132 <100 <10 <1,000 NA NA NA 133 <1,000 <1,000 >1,000 NA NA NA125 <100 <10 <1,000 <1,000 <10 <1,000 124 <100 <10 <1,000 <1,000 <10<1,000 127 <100 <10 >1,000 <1,000 <10 >1,000 126 <100 <10 <1,000 <1,000<10 <1,000 136 <1,000 <1,000 <100 NA NA NA 140 <100 <100 <100 NA NA NA141 <1,000 <1,000 <1,000 NA NA NA 142 <1,000 <1,000 <100 NA NA NA 143<100 <100 <10 NA NA NA 144 <100 <10 <10 NA NA NA 145 <1,000 <1,000<1,000 NA NA NA 147 <100 <1,000 >1,000 NA NA NA 150 >1,000 >1,000 >1,000NA NA NA 151 <1,000 <1,000 <1,000 NA NA NA 152 <1,000 <1,000 <100 NA NANA 153 <100 <100 <10 NA NA NA 154 <100 <10 <10 NA NA NA 155 >1,000<100 >1,000 NA NA NA 158 >1,000 <1,000 >1,000 NA NA NA 159 <1,000 <100<1,000 NA NA NA 163 >1,000 <10 <1,000 NA NA NA 162 >1,000 <100 <1,000 NANA NA 160 >1,000 <100 <1,000 NA NA NA 161 <1,000 <10 <1,000 NA NA NA 164<1,000 <10 <100 NA NA NA 165 <100 <10 <100 NA NA NA 166 <100 <10 >1,000NA NA NA 167 >1,000 >1,000 >1,000 NA NA NA 170 <1,000 <10 <1,000 NA NANA 171 <1,000 <10 <1,000 NA NA NA 168 <1,000 <100 >1,000 NA NA NA169 >1,000 <1,000 >1,000 NA NA NA 173 >1,000 >1,000 <100 NA NA NA175 >1,000 <1,000 <1,000 NA NA NA 174 <1,000 <100 <100 NA NA NA189 >1,000 >1,000 >1,000 NA NA NA 180 <1,000 <1,000 <1,000 NA NA NA 181<1,000 >1,000 <100 NA NA NA 183 <1,000 <1,000 <1,000 NA NA NA 182 >1,000<100 <1,000 NA NA NA 186 <1,000 <1,000 <1,000 NA NA NA 185 >1,000<1,000 >1,000 NA NA NA 184 <1,000 <1,000 <1,000 NA NA NA191 >1,000 >1,000 >1,000 NA NA NA 233 <100 <10 <10 <10 <10 <10 192<1,000 <10 <1,000 NA NA NA 193 <10 <10 <1,000 NA NA NA 194 <100 <10<1,000 NA NA NA 195 <1,000 <100 <1,000 NA NA NA 196 <100 <100 <10 NA NANA 197 <1,000 <1,000 <10 NA NA NA 201 <1,000 <10 <1,000 NA NA NA 206<1,000 <100 <1,000 NA NA NA 210 <100 <10 <1,000 NA NA NA 212 <1,000 <100<1,000 NA NA NA 213 <1,000 <1,000 <100 NA NA NA 218 >1,000 <10 <1,000 NANA NA 220 <1,000 <1,000 <100 NA NA NA 222 >1,000 >1,000 <1,000 NA NA NA227 >1,000 <1,000 <1,000 NA NA NA 224 >1,000 >1,000 >1,000 >1,000 >1,000<1,000 228 >1,000 >1,000 >1,000 NA NA NA 230 >1,000 <100 >1,000 NA NA NA176 >1,000 <1,000 <1,000 NA NA NA 178 <1,000 <1,000 <1,000 NA NA NA 179<1,000 >1,000 <1,000 NA NA NA 177 >1,000 >1,000 <1,000 NA NA NA225 >1,000 <10 <1,000 >1,000 >1,000 >1,000 232 <1,000 <1,000 <1,000 NANA NA

Example 32 Synthesis of(R)-3-Methanesulfonyloxymethyl-piperidine-1-carboxylic Acid Tert-ButylEster

To a stirred solution of N—BOC-3-piperidinemethanol (0.50 g, 0.002mmole) in DCM (10 mL) was added dropwise pyridine (2.42 mL, 0.03 mmole)followed by the addition of methanesulfonyl chloride (0.774 mL, 0.01mmole). The reaction mixture continued stirring at RT overnight. Thereaction mixture was quenched with water. The aqueous layer wasextracted with EtOAc (3×5 mL). Combined organic layers were dried overNa₂SO₄ and concentrated to yield a crude oil, 68, which was used in thenext step without further purification. LRMS: 193 (M+-BOC group).

Example 33 Synthesis of (R)-3-Phenoxymethyl-piperidine-1-carboxylic AcidTert-Butyl Ester

A solution of 68 (0.7 g, 2.4 mmole), Cs₂CO₃ (3.91 g, 12 mmole), andphenol (0.452 g, 4.8 mmole) in DMF (10 mL) was heated to 75° C. After 2h the reaction mixture was cooled down to RT and quenched with water.The aqueous layer was extracted with EtOAc (3×20 mL). Combined organiclayers were dried over Na₂SO₄ and concentrated to yield a crude oil.Chromatography with basic alumina (95:5 hexane:Et₂O) afforded 69 (196.6mg) as an oil. ¹H (300 MHz, CDCl₃) δ 7.33-7.27 (2H, m), 6.99-6.90 (3H,m), 3.86-3.80 (2H, m), 2.98-2.77 (4H, m), 2.0-1.20 (5H, M), 1.50 (s,9H).

Example 34 Synthesis of (R)-3-Phenoxymethyl-piperidine

Compound 69 (98.3 mg) was dissolved in DCM (2 mL) and cooled to 0° C. inan ice bath. TFA (0.8 mL) was added dropwise to the stirred cooledreaction mixture. After completion of addition the reaction continuedstirring at RT. Reaction progress was monitored by TLC and upon reactioncompletion reaction mixture was concentrated to yield the TFA salt of 70(98.3 mg). LRMS:M+192.

Example 35 Synthesis of(R)-[1-(4-Chloro-phenyl)-cyclobutyl]-(3-phenoxymethyl-piperidin-1-yl)-methadone

A solution of 70 (98.3 mg, 0.336 mmole),1-(4-chloro-phenyl)-cyclobutanecarboxylic acid (106.3 mg, 0.505 mmole),diisopropylethyl amine (0.234 mL, 1.34 mmole) in DCM (2 mL) was stirredat RT. PyBroP (235.4 mg, 0.505 mmole) was added and the reaction mixturecontinued stiffing at RT. After 6 h the reaction mixture was quenchedwith 10% KOH. Aqueous layer was extracted with EtOAc (3×2 mL). Combinedorganic layers were dried over Na₂SO₄ and concentrated to yield a crudeoil. Silica gel chromatography (4:1 hexane:EtOAc-1:4 hexane:EtOAc)afforded 71 (64.6 mg) as an oil. LRMS: M+384. ¹H (300 MHz, CDCl₃) δ7.39-7.29 (4H, m), 6.97 (2H, m), 6.90 (2H, m), 6.82 (1H, m), 4.58 (2H,d, J=12.5 Hz), 3.81-3.24 (4H, m), 2.97-1.161 (11H, m).

Example 36 Synthesis of(R)-[1-(4-Chloro-phenyl)-cyclobutyl]-3-phenoxymethyl-piperidine

A solution of 71 (62.8 mg, 0.16 mmole) in toluene (3 mL) was cooled to0° C. in an ice bath. RedAl (155.7 mg, 0.57 mmole) was added to thecooled reaction mixture. After completion of addition the reactionmixture continued stirring at RT. After 2 h the reaction mixture wasquenched with water. The aqueous layer was extracted with EtOAc (3×2mL). Combined organic layers were dried over Na₂SO₄ and concentrated toyield a crude oil. Silica gel:chromatography (9:1 hexane:Et₂O) afforded72 (52 mg) as an oil. LRMS:M+371.

Example 37 Synthesis of(R)-3#4-Trifluoromethyl)-phenoxymethyl-piperidine-1-carboxylic AcidTert-Butyl Ester

A solution of 68 (0.7 g, 2.4 mmole), Cs₂CO₃ (3.91 g, 12 mmole), and4-trifluoromethyl phenol (0.389 g, 2.4 mmole) in DMF (10 mL) was heatedto 75° C. After 4 h the reaction mixture was cooled down to RT andquenched with water. The aqueous layer was extracted with EtOAc (3×20mL). Combined organic layers were dried over Na₂SO₄ and concentrated toyield a crude oil. Chromatography with basic alumina (95:5 hexane:Et₂O)afforded 73 (70.3 mg) as an oil. LRMS:M+360.

Example 38 Synthesis of(R)-3-(4-Trifluoromethyl)-Phenoxymethyl-Piperidine

Compound 73 (66.9 mg) was dissolved in DCM (2 mL) and cooled to 0° C. inan ice bath. TFA (0.8 mL) was added dropwise to the stirred cooledreaction mixture. After completion of addition the reaction continuedstirring at RT. Reaction progress was monitored by TLC and upon reactioncompletion reaction mixture was concentrated to yield the TFA salt of 74(98.3 mg). LRMS:M+260.

Example 39 Synthesis of(R)-[1-(4-Chloro-phenyl)-cyclobutyl]-(3-(4-fluoromethyl)-phenoxymethyl-piperidin-1-yl)-methadone

A solution of 74 (98.3 mg, 0.362 mmole),1-(4-chloro-phenyl)-cyclobutanecarboxylic acid (114.3 mg, 0.543 mmole),diisopropylethyl amine (0.252 mL, 1.45 mmole) in DCM (2 mL) was stirredat RT. PyBroP (253.1 mg, 0.543 mmole) was added and the reaction mixturecontinued stirring at RT. After 12 h the reaction mixture was quenchedwith 10% KOH. Aqueous layer was extracted with EtOAc (3×2 mL). Combinedorganic layers were dried over Na₂SO₄ and concentrated to yield a crudeoil. Silica gel chromatography (100% hexane-100% EtOAc) afforded 75 (20mg) as an oil. LRMS:M+452.

Example 40 Synthesis of(R)-[1-(4-Chloro-phenyl)-cyclobutyl]-3-(4-trifluoromethyl)-phenoxymethyl-piperidine

A solution of 75 (450 mg, 0.996 mmole) in toluene (15 mL) was cooled to0° C. in an ice bath. RedAl (950.6 mg, 3.48 mmole) was added to thecooled reaction mixture. After completion of addition the reactionmixture continued stirring at RT. After 2 h the reaction mixture wasquenched with water. The aqueous layer was extracted with EtOAc (3×20mL). Combined organic layers were dried over Na₂SO₄ and concentrated toyield a crude oil. Silica gel:chromatography (85:15 hexane:EtOAc)afforded 76 (134 mg) as an oil. LRMS:M+437.

Example 41 Synthesis of[1-(4-Chloro-phenyl)-cyclobutyl]-[3(4-trifluoromethyl-phenylsulfanylmethyl)-piperidin-1-yl]-methanone

A solution of 43 (1.4 g, 3.6 mmole), Cs₂CO₃ (5.86 g, 18 mmole), and4-trifluoromethylthiol phenol (0.646 g, 3.6 mmole) in CH₃CN (50 mL) washeated to 75° C. After 4 h the reaction mixture was cooled down to RTand quenched with water. The aqueous layer was extracted with EtOAc(3×30 mL). Combined organic layers were dried over Na₂SO₄ andconcentrated to yield a crude oil. Chromatography using silica gel (4:1hexane:EtOAc) afforded 77 (196.6 mg) as an oil. LRMS: M+467.

Example 42 Synthesis of1-(4-Chloro-phenyl)-cyclobutyl]-[3(4-trifluoromethyl-phenylsulfanylmethyl)-piperidine

A solution of 77 (500 mg, 1.07 mmole) in toluene (13 mL) was cooled to0° C. in an ice bath. RedAl (750 mg, 3.74 mmole) was added to the cooledreaction mixture. After completion of addition the reaction mixturecontinued stirring at RT. After 1.5 h the reaction mixture was quenchedwith water. The aqueous layer was extracted with EtOAc (20 mL, 3×).Combined organic layers were dried over Na₂SO₄ and concentrated to yielda crude oil. Silica gel:chromatography (90:10 hexane:EtOAc) afforded 78(180 mg) as an oil. LRMS:M+452.

Example 43 Synthesis of2-(4-Chloro-phenyl)-1-(3-phenoxymethyl-piperidin-1-yl):propan-2-ol

A solution of 32 (0.343 mmol, 118 mg) in THF (1 mL) was treated withCH₃MgBr (3.0 M in ether) (3.0 equiv, 1.03 mmol, 343 μL) at 0° C. andstirred for 12 h. The reaction mixture was quenched with 10% HCl (5 mL)and then neutralized with NaHCO_(3(sat)) and extracted with EtOAc (2×10mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(PTLC, SiO₂, 20 cm×20 cm, 1 mm, 3:1 Hexane-EtOAc) provided 79 (29 mg,123 mg theoretical, 24%) as a colorless oil: R_(f) 0.44 (SiO₂, 3:1Hexane-EtOAc); LRMS m/z 361 (M⁺+1, C₂₁H₂₆ClNO₂, requires 361).

Example 44 Synthesis of{1-[2-(4-Chloro-phenyl)-2-methyl-propyl]-piperidin-3-yl]}-methanol

To amide 40 (100 mg, 0.34 mmol) in toluene (1 mL) was cautiously addedRed-Al (0.36 mL, 1.2 mmol). The resulting solution was allowed to stirat room temperature overnight. The reaction was then diluted with ethylacetate and quenched with 10% aqueous KOH. The layers were separated andthe aqueous layer further washed with ethyl acetate. The combinedorganic layers were then dried (MgSO₄), filtered, concentrated in vacuoand the resulting residue purified by flash column chromatography using4% 2M NH₃ in EtOH/DCM to provide the desired amine 80 (44 mg, 46%). LRMScalculated for C₁₆H₂₄ClNO 281.15, found (M+) 282.82. ¹H NMR (300 MHz,CDCl₃): 7.34 (d, J=8.7 Hz, 2H), 7.28 (d, J=8.7 Hz, 2H), 3.47-3.61 (m,2H), 2.47-2.52 (m, 1H), 2.42 (s, 2H), 2.12-2.29 (m, 4H), 1.38-1.70 (m,4H), 1.33 (s, 3H), 1.32 (s, 3H), 1.15-1.22 (m, 1H). ¹³C NMR (75 MHz,CDCl₃): 147.0, 131.4, 127.8, 127.6, 71.3, 67.1, 59.8, 56.6, 39.1, 37.5,27.0, 26.9, 26.6, 24.5.

Example 45 Synthesis of Methanesulfonic acid1-[1-(4-chloro-phenyl)-cyclobutanecarbonyl]-piperidin-3-ylmethyl Ester

To a suspension of lithium aluminum hydride (0.089 g, 2.34 mmol) inanhydrous tetrahydrofuran (25 mL) at 0° C. was added 44 (0.50 g, 1.17mmol). The reaction mixture was then stirred and refluxed for 6 h. Themixture was cooled to 0° C., and the reaction was quenched with slowaddition of water. The resulting salts were removed by vacuum filtrationthrough Celite, and the filtrate was partitioned between water anddiethyl ether (50 mL each). The aqueous layer was extracted well withdiethyl ether (4×50 mL), and the combined organic portions were driedover anhydrous magnesium sulfate, filtered and concentrated by rotaryevaporation. The organic residue was purified by flash chromatographeyon silica gel, eluting with dichloromethane/2.0 M ammonia in ethylalcohol (96:4) to give 83 (0.15 g, 31%) as a pale yellow gum;C₂₄H₂₈ClNO₃, LRMS (m/z)=414 (MH+).

Example 46 Synthesis of[1-(4-Chlorophenyl)-cyclobutyl]-[3-(4-fluorophenoxymethyl)-piperidin-1-yl]-methanone(84)

Compound 84 was synthesized from 4-fluorophenol (0.29 g, 2.60 mmol),cesium carbonate (1.27 g, 3.89 mmol) and compound 43 (1.0 g, 2.60 mmol),using the method described for the synthesis of compound 44 to give 0.60g of the desired product 84. C₂₃H₂₅ClFNO₂, LRMS (m/z)=402 (MH+).

Example 47 Synthesis of[1-(4-Chloro-phenyl)-cyclobutyl]-[3-(pyridin-3-yloxymethyl)-piperidin-1-yl]-methanone(85)

Compound 85 was synthesized from 3-hydroxypyridine (0.25 g, 2.60 mmol),cesium carbonate (1.27 g, 3.89 mmol) and compound 43 (1.0 g, 2.60 mmol),using the method described for the synthesis of compound 44 to give 0.52g of the desired product 85. C₂₂H₂₅ClN₂O₂, LRMS (m/z)=385 MH+).

Example 48 Synthesis of1-[1-(4-Chloro-phenyl)cyclobutylmethyl]-3-(4-fluoro-phenoxymethyl)-piperidine(86)

Compound 86 was synthesized from compound 84 (0.50 g, 1.25 mmol) andlithium aluminum hydride (0.10 g), using the method described for thesynthesis of compound 83 to give 0.24 g of the desired product 86.C₂₃H₂₇ClFNO, LRMS (m/z)=388 (MH+).

Example 49 Synthesis of3-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-ylmethoxy}-pyridine

Compound 87 was synthesized from compound 85 (0.50 g, 1.25 mmol) andlithium aluminum hydride (0.10 g), using the method described for thesynthesis of compound 83 to give 0.19 g of the desired product 87.C₂₂H₂₇ClN₂O, LRMS (m/z)=371 (MH+).

Example 50 Synthesis of1-[1-(4-Chlorophenyl)cyclobutylmethyl]piperidin-3-ol (89)

The synthesis of amide 88 from commercially-available3-hydroxypiperidine hydrochloride and1-(4-chloro-phenyl)-cyclobutanecarboxylic acid is described in Example72.

To amide 88 (100 mg, 0.34 mmol) in toluene (1 mL) was cautiously addedRed-Al (0.36 mL, 1.2 mmol). The resulting solution was allowed to stirat room temperature for one and one half hours before diluting withethyl acetate and quenching with 10% aqueous KOH. The layers wereseparated and the aqueous layer further washed with ethyl acetate. Thecombined organic layers were then dried (MgSO₄), filtered andconcentrated in vacuo. The resulting residue was purified by flashcolumn chromatography using 4% 2M NH₃ in EtOH/DCM to provide the desiredamine 89 (51 mg, 54%). LRMS calculated for C₁₆H₂₂ClNO 279.14, found (M+)280.87. ¹H NMR (300 MHz, CDCl₃): 7.28 (d, J=8.3 Hz, 2H), 7.11 (d, J=8.3Hz, 2H), 3.67-3.69 (m, 1H), 2.73 (m, 1H), 2.66 (s, 2H), 2.14-2.44 (m,7H), 1.97-2.11 (m, 2H), 1.79-1.90 (m, 1H), 1.24-1.66 (m, 4H). ¹³C NMR(75 MHz, CDCl₃): 147.6, 131.1, 127.9, 127.4, 68.2, 66.0, 62.2, 55.3,46.9, 31.8, 31.7, 30.8, 21.2, 16.0.

Example 51 Synthesis of3-[1-(4-Chloro-phenyl)-cyclobutanecarbonyl]-cyclohexanecarbaldehyde

To a stirred solution of pyridinium chlorochromate (210 mg, 0.98 mmole)in anhydrous dichloromethane (5 mL) was added alcohol 1 (200 mg, 0.65mmole) dissolved in anhydrous dichloromethane (5 mL). After completionof addition the reaction mixture continued stirring at RT for 5 h. Thereaction mixture was then filtered though a presaturated silica gel plug(1:1 hexane:EtOAc) to obtain the aldehyde 90 as a clear oil (100 mg,50%).

Example 52 Synthesis of3-[1-(4-Chloro-phenyl)-cyclobutyl]-{3-{2-(4-trifluoromethyl-phenyl)-vinyl]-cyclohexyl}-methanone

To a stirring solution of triphenylphosphine (2.2 g, 8.4 mmole) inahydrous diethylether (8 mL) was added 4-trifluoromethyl-benzyl bromide(2 g, 8.4 mmole) dissolved in ahydrous diethylether (7 mL). The reactionmixture continued stirring at RT for 72 h. The phosphine salt, B, wascollected via filtration of the reaction mixture. The salt, white solid,was dried under vacuum (2.52 g).

A solution of B (2.5 mmol, 754 mg) in THF (10 mL) was treated with nBuLi(1.6M in hexanes, 3.7 mmole, 2.3 mL) at −78° C. The solution was warmedto 0° C. for 30 min and then cooled again to −78° C. A solution of 90(3.7 mmol, 1.85 g) in THF (10 mL) was added to the above reactionmixture at −78° C. The reaction was stirred for 2 h. The reactionmixture was quenched with water and then extracted with EtOAc (3×20 mL).The combined organics were dried over Na₂SO_(4(s)). The solvents wereremoved in vacuo and the crude material was purified using silica gelchromatography (100% hexanes-85:15 hexanes:EtOAc) to provide 91 (176 mg,17%) as an oil. LRMS: M+415.

Example 53 Synthesis of(1-phenyl-cyclobutyl]-{3-{2-(4-trifluoromethyl-phenyl)-ethyl]-cyclohexyl}-methanone

A solution of 91 (592 mmol, 190 mg) in CH₃OH (5 mL) was treated with 10%Pd—C (60 mg) and H₂ (Pair Hydrogenator, 65 psi). The reaction was shakenfor 4 h. The reaction mixture was filtered, and the solvents wereremoved in vacuo to provide 92 (180 mg). ¹H (300 MHz, CDCl₃) δ 7.55 (2H,t, J=9 Hz), 7.38-7.29 (m, 3H), 7.24 (2H, d, J=7.3 Hz), 7.14 (2H, d, J=9Hz), 4.64 (2H, m), 4.42 (2H, m), 3.80-0.78 (m, 15H).

Example 54 Synthesis of1-(1-phenyl-cyclobutylmethyl-3-[2-(4-trifluoromethyl-phenyl)-ethyl]-piperidine

A solution of amide 92 (180 mg, 0.50 mmole) in anhydrous toluene (5 mL)was cooled to 0° C. RedAl (356 mg, 1.76 mmole) was added to the cooledstirring reaction mixture. After completion of addition, the reactioncontinued stirring at RT. After 2 h, the reaction mixture was dilutedwith EtOAc and quenched with water. The aqueous layer was extracted withEtOAc (3×10 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated to yield an oil. The crude material was purified usingsilica gel chromatography (100% hexane-80:20 Hexanes:EtOAc) to yield 93.LRMS: M+402.

Example 55 Synthesis of1-(4-Chloro-phenyl)-cyclobutyl]-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-methanone

The olefin 91 (100 mg, 0.28 mmole) was dissolved in anhydrousdichloromethane (3 mL) and placed in a two-neck flask. To this solutionplatinum oxide (16 mg, 0.07 mmole) was added. The system was alternatelyevacuated and filled with nitrogen, then hydrogen from a balloon. Thereaction mixture was stirred vigorously under hydrogen for 4 h. Thecrude solution was filtered and concentrated to yield 94 as a milkywhite oil (97 mg, 77%). ¹H (300 MHz, CDCl₃) δ 7.55 (2H, d, J=8.3 Hz),7.34-7.31 (m, 4H), 7.17 (2H, d, J=7.6 Hz), 4.62 (2H, m), 4.41 (2H, m),3.00-0.85 (m, 15H).

Example 56 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-[2-(4-trifluoromethyl-phenyl)-ethyl]-piperidine

A solution of amide 94 (97 mg, 0.27 mmole) in anhydrous toluene (2 mL)was cooled to 0° C. RedAl (192 mg, 0.94 mmole) was added to the cooledstirring reaction mixture. After completion of addition, the reactioncontinued stirring at RT. After 2 h, the reaction mixture was dilutedwith EtOAc (4 mL) and quenched with water. The aqueous layer wasextracted with EtOAc (3×5 mL). The combined organic layers were driedover Na₂SO₄ and concentrated to yield an oil. The crude material waspurified using silica gel chromatography (100% hexane-80:20:0.2%hexanes:EtOAc: 2M NH₃ in EtOH) to yield 95. LRMS: M+435.

Example 57 Synthesis of 3-(1-hydroxy-ethyl)-piperidine-1-carboxylic acid2,2-dimethyl-propyl Ester

To a cooled solution of N—BOC-piperidine-3-carboxaldehyde dissolved indiethyl ether (50 mL) was added the methyl grignard reagant (10.55 mL,10.5 mmol, 1 M in diethyl ether). After completion of addition thereaction mixture continued stirring at 0° C. for 15 min. and was thenwarmed to RT. After 15 min. of stirring at RT the reaction mixture wasquenched with water. The aqueous layer was extracted with EtOAc (3×100mL). The combined organic layer was then dried over Na₂SO₄, filtered andconcentrated in vacuo. The resulting residue was purified by silica gelchromatography using a gradient (100% hexanes-1:1 hexanes:EtOAc) toobtain the desired alcohol 96. ¹H NMR (300 MHz, CDCl₃): δ 3.94 (m, 1H),3.65 (t, 1H, J=6.1 Hz), 2.73 (m, 2H), 2.65 (m, 2H), 1.94 (m, 2H), 1.70(m, 2H), 1.50 (s, 9H), 1.29 (m, 3H).

Example 58 Synthesis of[1-(4-chlorophenyl)-cyclobutyl]-[3-(1-hydroxy-ethyl)-piperidin-1-yl]-methanone

To a cooled solution of the protected amine 96 (750 mg, 3.3 mmol)dissolved in DCM (4 mL) was added concentrated TFA (4 mL) dropwise.After completion of addition the reaction continued stirring at 0° C.After 1.5 h the reaction mixture was concentrated in vacuo to yield theTFA salt as a brown oil.

This salt was dissolved in DCM (16.5 mL) and solid1-(4-chloro-phenyl)cyclobutane carboxylic acid (1.43 g, 4.95 mmole)followed by di-isopropyl ethyl amine (2.3 mL 13.2 mmol) were added.After completion of addition solid PyBroP (2.29 g, 4.95 mmole) was addedto the stirring reaction mixture. The reaction mixture continuedstirring at RT for 10 h and was quenched with water and 10% KOH. Theaqueous layer was extracted with EtOAc (3×20 mL). Combined organiclayers were dried over Na₂SO₄ and concentrated to yield an oil. Thiscrude material was purified using silica gel chromatography (1:1hexane:EtOAc) to yield 97 as an oil. LRMS: M+321.

Example 59 Synthesis of1-[1-(4-chloro-phenyl)-cyclobutyl]-{3-[1-(4-trifluoromethyl-phenoxy)-ethyl]-piperidin-1-yl}-methanone

A solution of 97 (300 mg, 0.932 mmoles), triphenylphosphine (370 mg,1.40 mmole), and phenol (300 mg, 1.86 mmoles) dissolved in anhydrousether (2.5 mL) was cooled in a brine bath to −5° C. DEAD (240 mg, 1.40mmoles) dissolved in ether (2.5 mL) was added to the cooled stirringreaction mixture. After completion of addition the reaction mixturecontinued stirring at −5° C. After 4 h the reaction mixture wasconcentrated and crude material was dissolved in a hexane/ethyl acetatemixture (70% hexanes:30% ethyl acetate, 30 mL). Phosphine by-productsprecipitated and were filtered off. Filtrate was concentrated to yieldan oil. This oil was purified using silica gel chromatography (3:2hexanes:EtOAc-100% EtOAc) to yield 98. LRMS: M+367.

Example 60 Synthesis of1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-3-[1-(4-trifluoromethyl-phenoxy)-ethyl]-piperidine

A solution of 98 (215 mg, 0.461 mmoles) dissolved in anhydrous toluene(2.3 mL) was cooled to 0° C. RedAl (326 mg, 1.62 mmoles) was addeddropwise to the cooled stirring reaction mixture. After completion ofaddition the reaction continued stirring at RT. After 1.5 h water wasadded to the reaction mixture. The aqueous layer was extracted withEtOAc 3× (5 mL). Combined organic layers were dried over Na₂SO₄ andconcentrated to yield an oil. The crude material was purified usingsilica gel chromatography (1:1 Hexanes:EtOAc) to yield 99. ¹H NMR (300MHz, CDCl₃): δ 7.54 (dd, 2H, J=8.7 Hz), 7.27 (d, 2H, J=6.2 Hz), 7.10 (m,2H), 6.86 (d, 2H, J=8.5 Hz), 4.10 (m, 1H), 2.69-1.08 (m, 20H).

Example 61 Synthesis of 2-benzyl Aminoethanol

To a stirring solution of benzaldehyde (41.60 g, 393 mmole) dissolved inanhydrous MeOH (350 mL) was added 2-amino ethanol (20 g, 327 mmole)dropwise. After completion of addition the reaction mixture was heatedto 75° C. After 0.5 h the reaction mixture was cooled to RT and placedin an ice bath. Solid NaBH₄ (18.58 g, 491 mmole) was added over 20 min.After completion of addition the reaction mixture continued stirring atRT. After 10 h the reaction mixture was concentrated and the white crudematerial was taken up in DCM (300 mL). The organic layer was extractedwith water (1×200 mL). The aqueous layer was acidified with 10% HCl andthen extracted with DCM (3×200 mL). Combined organic layers were driedover Na₂SO₄ and concentrated to yield 2-benzyl aminoethanol (55.21 g,0.363 moles, 92%). LRMS: M+ 152.

Example 62 Synthesis of 4-benzyl-2-chloromethyl-morpholine

A solution of 2-benzyl aminoethanol (7.0 g, 46.3 mmoles) andepichlorohydrin (42.8 g, 463 mmoles) was heated to 40° C. After 2.5 hthe reaction was cooled to RT and the excess epichlorohydrin wasevaporated in vacuo. Sulfuric acid (14 mL) was added slowly to the crudemixture. After completion of addition the reaction flask was placed in apreheated oil bath (150° C.). The reaction mixture was heated for 30minutes, cooled to RT, and quenched with ice. The aqueous layer wasbasified with 10% KOH and extracted with EtOAc 3× (300 mL). Combinedorganic layers were dried over Na₂SO₄ and concentrated to yield a crudeoil. This oil was purified using silica gel chromatography (90:8:2hexanes:DCM: 2M NH₃ in EtOH) to obtain the morpholine (3.41 g, 15.16mmole, 33%). ¹³C NMR (100 MHz, CDCl₃) δ 137.7, 129.4, 128.6, 127.6,75.4, 67.1, 63.4, 56.1, 53.0, 45.2. LRMS: 225.

Example 63 Synthesis of 2 chloromethyl-morpholine

4-Benzyl-2-chloromethyl-morpholine (316 mg, 1.40 mmole) dissolved inacetic acid (3.16 mL) was hydrogenated in the presence of palladium oncharcoal (10%, 94.8 mg) under pressure (50 psi) at RT. After 5 h thereaction catalyst was removed by filtration and the filtrate wasconcentrated to yield 2-chloromethyl-morpholine as an oil. ¹³C NMR (100MHz, CDCl₃) δ 74.6, 66.5, 47.0, 44.7, 44.1. LRMS: M+136.

Example 64 Synthesis of(2-chloromethyl-morpholin-4-yl)-[1-(4-chloro-phenyl)-cyclobutyl]-methanone

To a solution of 2-chloromethyl-morpholine (163 mg, 1.2 mmole) and EDCI(280 mg, 1.8 mmole) dissolved in DCM (5 mL) was added solid1-(4-chloro-phenyl)-cyclobutane carboxylic acid (304 mg, 1.44 mmole)followed by di-isopropyl ethyl amine (310 mg, 2.4 mmol). The reactionmixture continued stirring at RT for 10 h and was quenched with water.The aqueous layer was extracted with EtOAc (3×10 mL). Combined organiclayers were dried over Na₂SO₄ and concentrated to yield an oil. Thiscrude material was purified using silica gel:chromatography (1:1hexane:EtOAc) to yield 100 (100 mg, 12.7%). LRMS: M+ 328.

Example 65 Synthesis of[1-(4-chloro-phenyl)-cyclobutyl]-[2-(4-trifluoromethyl-phenoxymethyl)-morpholin-4-yl]methanone

To a solution of KOH (34 mg, 0.61 mmoles) dissolved in DMSO (1.5 mL) wasadded the phenol (49 mg, 0.30 mmoles) followed by the halide 100 (100mg, 0.30 mmoles). After completion of addition the reaction mixture washeated to 55° C. After 12 h the reaction mixture was cooled to RT andquenched with water. The aqueous layer was extracted with EtOAc 3× (2mL) and combined organic layers were dried over Na₂SO₄ and concentratedto yield a crude oil. The crude material was purified using silica gelchromatography (1:1 hexane:EtOAc) to yield 101 (16 mg, 0.035 mmole,12.3%). LRMS: M+ 353.

Example 66 Synthesis of[1-(4-chloro-phenyl)-cyclobutyl]-[2-(4-trifluoromethyl-phenoxymethyl)-morpholin-4-yl]methanone

To a cooled solution of LAH (0.053 mL, 0.053 mmol, 1 M solution in THF)in anhydrous THF was added 101 (16 g, 0.035 mmole) dissolved inanhydrous THF (0.175 mL). After completion of addition the reactioncontinued stirring at RT. After 4 h the reaction mixture was quenchedwith 5% HCl (aq.). The aqueous layer was extracted with EtOAc (3×2 mL)and combined organic layers were dried over Na₂SO₄ and then concentratedto yield an oil. The crude material was purified using a silica gel prepplate (90:10 Hexanes:EtOAc) to yield 102. LRMS: M+440.

Example 67 Synthesis of azetidine-3-carboxylic Acid Ethyl EsterHydrochloride

Hydrogen chloride gas was gently bubbled into a solution ofazetidine-3-carboxylic acid (1 g, 9.85 mmole) in methanol (20 mL). After3 min. the HCl gas source was removed from the solution and the reactionflask was capped. The reaction mixture continued stirring at RT. After 2days the reaction mixture was concentrated in vacuo to yield the HClsalt as a yellow oil (762 mg, 4.60 mmole, 47%). ¹³C NMR (100 MHz, CDCl₃)δ 171.1, 53.2, 48.2, 34.7.

Example 68 Synthesis of1-[1-(4-chloro-phenyl)-cyclobutanecarbonyl]-azetidine-3-carboxylic AcidMethyl Ester

To a stirred solution of azetidine-3-carboxylic acid methyl esterhydrochloride (762 mg, 4.60 mmoles) and 1-(4-chloro-phenyl)-cyclobutanecarboxylic acid (2.09 g, 9.90 mmoles) in anhydrous DCM (20 mL) was addeddi-isopropyl ethyl amine (4.6 mL, 26.4 mmoles) dropwise. Aftercompletion of addition solid PyBroP (4.63 g, 9.94 mmoles) was added tothe stirring reaction mixture. The reaction mixture continued stirringat RT for 10 h and was quenched with water. The aqueous layer wasextracted with EtOAc (3×20 mL). Combined organic layers were dried overNa₂SO₄ and concentrated to yield an oil. This crude material waspurified using silica gel chromatography (9:1 hexanes:EtOAc-1:1hexane:EtOAc) to yield the desired amide (1.0 g, 65%). ¹H NMR (300 MHz,CDCl₃): δ 7.38-7.29 (m, 4H), 4.16 (m, 2H), 3.88 (m, 1H), 3.72 (s, 3H),3.31-1.63 (m, 8H). LRMS: M+309.

Example 69 Synthesis of{1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-azetidin-3-yl}-methanol

To a cooled (0° C.) solution of LAH (9.8 mL, 9.8 mmol, 1 M solution inTHF) in anhydrous THF was added the ester (1 g, 3.0 mmole) dissolved inanhydrous THF (20 mL). After completion of addition the reactioncontinued stirring at RT. After 3 h the reaction mixture was quenchedwith 10% HCl (aq.). The aqueous layer was extracted with EtOAc (3×200mL) and combined organic layers were dried over Na₂SO₄ and thenconcentrated to yield 103 as an oil. LRMS: M+265.

Example 70 Synthesis of Methanesulfonic Acid1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-azetidin-3-ylmethyl Ester

To a solution of primary alcohol 103 (1.8 g, 6.77 mmol) in DCM (30 mL)at room temperature was added iPr₂NEt (3 mL, 16.93 mmol) followed byMsCl (0.6 mL, 7.44 mmol). The reaction mixture was allowed to stir forone hour before purifying the crude mixture using a presaturated silicagel plug (4:1 hexanes:EtOAc) to provide the desired mesylate 104, whichwas used in the next reaction without further purification. LRMS: M+344.

Example 71 Synthesis of Methanesulfonic Acid1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-azetidine

To the mesylate 104 (2.3 g, 6.70 mmol) in anhydrous acetonitrile (33 mL)was added α,α,α-trifluoro-p-cresol (1.1 g, 6.70 mmol) followed by Cs₂CO₃(10.90 g, 33.44 mmol). The reaction mixture was heated to 75° C. and thereaction progress was monitored by HPLC. Upon completion the reactionmixture was quenched with water and the aqueous layer was extracted withEtOAc (3×20 mL). The combined organic layer was then dried over Na₂SO₄,filtered and concentrated in vacuo. The resulting residue was purifiedby silica gel chromatography using a gradient (30% hexanes:70% EtOAc) toobtain the desired ether 105. LRMS: M+410.

Example 72 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-benzyloxy)-piperidine

Amide 88 was prepared from commercially-available 3-hydroxypiperidinehydrochloride and 1-(4-chloro-phenyl)cyclobutane carboxylic acid, usingthe procedure outlined for the synthesis of 1 in Example 1:3-hydroxypiperidine hydrochloride (1.0 g, 7.29 mmol),1-(4-chlorophenyl)-1-cyclobutane carboxylic acid (2.29 g, 10.9 mmol),PyBroP (5.08 g, 10.9 mmol), iPr₂NEt (6.33 mL, 36.3 mmol), DCM (40 mL).Purification by flash column chromatography using 40% ethylacetate/petroleum ether provided the desired amide 88 (1.74 g, 82%). ¹³CNMR (75 MHz, CDCl₃): 6 (major rotamer only) 174.4, 141.9, 132.1, 128.9,126.4, 65.8, 51.9, 49.3, 45.5, 32.6, 31.9, 22.0, 15.2.

To amide 88 (200 mg, 0.68 mmol) in DMF (3 mL) was cautiously addedsodium hydride (82 mg, 2.0 mmol). The resulting mixture was allowed tostir at room temperature for 45 minutes before adding4-(trifluoromethyl)benzoyl bromide (179 mg, 0.75 mmol). The reaction wasallowed to continue stirring at room temperature overnight beforediluting with ethyl acetate and quenching with a saturated aqueoussodium chloride solution. The layers were separated and the aqueouslayer further washed with ethyl acetate. The combined organic layerswere then dried (MgSO₄), filtered, concentrated in vacuo and theresulting residue purified by flash column chromatography using 30%ethyl acetate/hexane to provide the desired ether 106 (192 mg, 62%).

Amide 106 was reduced as per the procedure for the reduction of 88, seeExample 50: 106 (91 mg, 0.202 mmol), Red-Al (0.212 mL, 0.706 mmol),toluene (1 mL). Purification by flash column chromatography using 1% 2MNH₃ in EtOH/DCM provided the desired amine 107. LRMS calculated forC₂₄H₂₇ClF₃NO 437.17, found 437.71. ¹H NMR (300 MHz, CDCl₃): δ 7.58 (d,J=8.0 Hz, 2H), 7.36 (d, J=7.9 Hz, 2H), 7.22 (d, J=8.3 Hz, 2H), 7.06 (d,J=7.9 Hz, 2H), 4.36 (s, 2H), 3.2 (m, 1H), 2.66-2.71 (m, 1H), 2.50-2.55(m, 1H), 2.35-2.37 (m, 2H), 1.97-2.26 (m, 6H), 1.77-1.94 (m, 3H),1.53-1.58 (m, 1H), 1.29-1.44 (m, 1H), 1.04-1.18 (m, 1H).

Example 73 Synthesis of(R)-3-(Benzo[1,3]-dioxol-5-yloxymethyl)-1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-piperidine

Amide 108 was prepared from commercially-available (R)-ethyl nipecotateL-tartrate and 1-(4-chlorophenyl)-1-cyclobutane carboxylic acid, usingthe procedure outlined for the synthesis of 1 in Example 1: (R)-ethylnipecotate L-tartrate (10.0 g, 32.6 mmol),1-(4-chlorophenyl)-1-cyclobutane carboxylic acid (10.3 g, 48.9 mmol),PyBroP (22.8 g, 48.9 mmol), iPr₂NEt (28.0 mL, 163 mmol), DCM (170 mL).Purification by flash column chromatography using 35% ethylacetate/hexane provided the desired amide 108 (5.1 g, 45%).

To a flask containing LiAlH₄ (1.66 g, 43.8 mmol), charged with Argon at0° C. was added tetrahydrofuran (50 mL). After the addition wascomplete, the suspension was allowed to warm to room temperature forfive minutes before recooling to 0° C. Next, a solution of amide 108(5.1 g, 14.6 mmol) in tetrahydrofuran (25 mL) was added over fiveminutes. After continuing at this temperature for fifteen minutes, thereaction was allowed to warm to room temperature and stir overnightbefore recooling to 0° C. and cautiously quenching by the addition of 1NH₂SO₄. The aqueous layer was then basified by the addition of saturatedaqueous NaHCO₃. The resulting mixture was filtered through a pad ofcelite, washing with ethyl acetate. The layers were separated and theaqueous layer further washed with ethyl acetate. The combined organiclayers were dried (MgSO₄), filtered, concentrated in vacuo and theresulting residue purified by flash column chromatography using agradient of 2 to 4% 2M NH₃ in EtOH/DCM to provide the desired 109 (2.87g, 67%). ¹H NMR (300 MHz, CDCl₃): δ 7.26-7.29 (m, 2H), 7.12-7.16 (m,2H), 3.58-3.64 (m, 1H), 3.47-3.53 (m, 1H), 3.00 (m, 1H), 2.64 (s, 2H),2.43-2.48 (m, 1H), 2.15-2.32 (m, 7H), 1.99-2.09 (m, 1H), 1.79-1.92 (m,1H), 1.36-1.69 (m, 4H), 1.16-1.29 (m, 1H). ¹³C NMR (75 MHz, CDCl₃): δ147.9, 131.0, 127.9, 127.4, 69.1, 67.6, 59.7, 56.2, 46.7, 37.0, 31.8,31.5, 27.2, 24.3, 15.9.

Alcohol 109 could be converted to the desired 110 using the procedureoutlined for the conversion of 36 to 37 in Example 22 with thesubstitution of sesamol for α,α,α-trifluoro-p-cresol. Mesylateformation: 109 (2.82 g, 9.61 mmol), iPr₂NEt (4.18 mL, 24.0 mmol), MsCl(0.818 mL, 10.6 mmol), DCM (44 mL). After purification by flash columnchromatography using 2% 2M NH₃ in EtOH/DCM the desired mesylate wasprovided (3.46 g, 97%). Mesylate displacement: mesylate (3.46 g, 9.31mmol), Cs₂CO₃ (7.59 g, 23.4 mmol), sesamol (1.29 g, 9.31 mmol), DMF (50mL). Purification by flash column chromatography using 1% 2M NH₃ inEtOH/DCM followed by a second column using a gradient of 10 to 20% ethylacetate/hexane provided the desired 110. LRMS calculated for C₂₄H₂₈ClNO₃413.18, found (M+) 414.27. ¹H NMR (300 MHz, CDCl₃): δ 7.21 (d, J=8.5 Hz,2H), 7.07 (d, J=8.4 Hz, 2H), 6.69 (d, J=8.4 Hz, 1H), 6.43 (d, J=2.4 Hz,1H), 6.23 (dd, J=2.4, 8.4 Hz, 1H), 5.89 (s, 2H), 3.53-3.65 (m, 2H),2.44-2.57 (m, 3H), 2.16-2.31 (m, 5H), 1.79-2.08 (m, 5H), 1.56-1.62 (m,1H), 1.42-1.48 (m, 2H), 0.95-1.09 (m, 1H). ¹³C NMR (75 MHz, CDCl₃): δ154.6, 148.3, 148.1, 141.3, 130.7, 127.7, 127.5, 107.8, 105.4, 101.0,97.9, 71.4, 68.7, 58.9, 56.3, 47.0, 36.1, 31.6, 26.7, 24.6, 16.0. eedetermination: 96.4%

The HCl salt of 110 could be prepared by dissolving the basic amine inacetonitrile and adding an excess of 2M HCl. The acetonitrile could thenbe removed in vacuo and the sample frozen and liopholized to provide thedesired salt as a white solid. LRMS calculated for C₂₄H₂₈ClNO₃ (freebase) 413.18, found 413.88. ¹H NMR (300 MHz, CDCl₃): δ 7.31-7.39 (m,4H), 6.63 (d, J=8.4 Hz, 1H), 6.31 (d, J=2.0 Hz, 1H), 6.11-6.15 (m, 1H),5.86 (s, 2H), 3.66 (dd, J=3.8, 9.5 Hz, 1H), 3.48-3.54 (m, 3H), 3.08-3.22(m, 2H), 2.76-2.91 (m, 1H), 2.20-2.64 (m, 7H), 2.05-2.15 (m, 1H),1.84-1.94 (m, 1H), 1.63-1.75 (m, 2H), 1.15-1.28 (m, 1H). ¹³C NMR (75MHz, CDCl₃): δ 153.7, 148.2, 144.1, 141.9, 132.8, 129.2, 127.8, 107.8,105.3, 101.1, 97.9, 70.1, 68.0, 57.4, 54.9, 44.2, 33.3, 32.8, 24.6,21.8, 15.8. [α]=−6.1 (c=0.74, MeOH).

Example 74 Synthesis of(S)-3-(Benzo[1,3]dioxol-5-yloxymethyl)-1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-piperidine

113 was prepared from ethyl (S)-nipecotate D-tartrate and1-(4-chloro-phenyl)-cyclobutanecarboxylic acid, using the procedureoutlined in Example 73 for the synthesis of 110.

Preparation of 111: (S)-ethyl nipecotate L-tartrate (10.3 g, 33.6 mmol),1-(4 chlorophenyl)-1-cyclobutane carboxylic acid (10.6 g, 50.4 mmol),PyBroP (23.5 g, 50.4 mmol), iPr₂NEt (29.3 mL, 168 mmol), DCM (170 mL).Purification by flash column chromatography using 35% ethylacetate/hexane provided the desired amide 111 (5.5 g, 47%).

Preparation of 112: 111 (5.50 g, 15.8 mmol), LiAlH₄ (1.79 g, 47.3 mmol),THF (75 mL). Purification by flash column chromatography using agradient of 2 to 4% 2M NH₃ in EtOH/DCM provided the desired 112 (3.79 g,82%).

Preparation of 113. Mesylate formation: amino alcohol (3.79 g, 12.9mmol), iPr₂NEt (5.63 mL, 32.3 mmol), MsCl (1.10 mL, 14.2 mmol), DCM (60mL). After purification by flash column chromatography using 2% 2M NH₃in EtOH/DCM the desired mesylate was provided (4.73 g, 98%). Mesylatedisplacement: mesylate (4.73 g, 12.7 mmol), Cs₂CO₃ (10.3 g, 31.8 mmol),sesamol (1.76 g, 12.7 mmol), DMF (65 mL). Purification by flash columnchromatography using a gradient of 5 to 10% ethyl acetate/hexanefollowed by a second column using 1% 2M NH₃ in EtOH/DCM provided thedesired 113 (3.52 g, 67%). LRMS calculated for C₂₄H₂₈ClNO₃ 413.18, found413.73. ¹H NMR (300 MHz, CDCl₃): δ 7.20 (d, J=8.6 Hz, 2H), 7.07 (d,J=8.3 Hz, 2H), 6.69 (d, J=8.5 Hz, 1H), 6.43 (d, J=2.4 Hz, 1H), 6.23 (dd,J=2.4, 8.4 Hz, 1H), 5.89 (s, 2H), 3.54-3.65 (m, 2H), 2.34-2.57 (m, 3H),2.16-2.31 (m, 5H), 1.74-2.08 (m, 5H), 1.56-1.62 (m, 1H), 1.35-1.51 (m,2H), 0.97-1.09 (m, 1H). ¹³C NMR (75 MHz, CDCl₃): δ 154.6, 148.3, 148.1,141.3, 130.7, 127.7, 127.5, 107.8, 105.4, 101.0, 97.9, 71.5, 68.7, 58.9,56.3, 47.0, 36.1, 31.6, 26.7, 24.6, 16.0. ee determination: 98.7% TheHCl salt of 113 could be prepared by dissolving the basic amine inacetonitrile and adding an excess of 2 M HCl. The acetonitrile couldthen be removed in vacuo and the sample frozen and lypholized to providethe desired salt as a white solid. LRMS calculated for C₂₄H₂₈ClNO₃ (freebase) 413.18, found 413.44. ¹H NMR (300 MHz, CDCl₃): δ 7.32-7.37 (m,4H), 6.62 (d, J=8.4 Hz, 1H), 6.31 (d, J=2.4 Hz, 1H), 6.11-6.15 (m, 1H),5.86 (s, 2H), 3.64-3.69 (m, 1H), 3.49-3.54 (m, 3H), 3.10-3.23 (m, 2H),2.81 (m, 1H), 2.26-2.61 (m, 7H), 2.03-2.14 (m, 1H), 1.83-1.93 (m, 1H),1.63-1.74 (m, 2H), 1.14-1.27 (m, 1H). ¹³C NMR (75 MHz, CDCl₃): δ. 153.7,148.2, 144.0, 141.9, 132.8, 129.2, 127.8, 107.8, 105.3, 101.1, 97.9,70.1, 68.0, 57.4, 54.9, 44.2, 33.4, 32.8, 24.6, 21.8, 15.8. [α]=+5.4(c=0.78, MeOH).

Example 75 Synthesis of(g)-1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)piperidine

Amide 108 was prepared from commercially-available (R)-ethyl nipecotateL-tartrate and 1-(4-chlorophenyl)-1-cyclobutane carboxylic acid, usingthe procedure outlined for the synthesis of 1 in Example 1: (R)-ethylnipecotate L-tartrate (17.0 g, 55 mmol),1-(4-chlorophenyl)-1-cyclobutane carboxylic acid (17.43 g, 83 mmol),PyBroP (38.51 g, 83 mmol), iPr₂NEt (68 mL), DCM (240 mL). Purificationby flash column chromatography using 10% ethyl acetate/hexane providedthe desired amide 108 (9.56 g, 44%).

To a flask containing LiAlH₄ (2.75 g, 72.0 mmol), charged with Argon at0° C. was added tetrahydrofuran (50 mL). After the addition wascomplete, the suspension was cooled to 0° C. Next, a solution of amide108 (9.56 g, 24 mmol) in tetrahydrofuran (50 mL) was added dropwise.After completion of addition, the reaction was allowed to warm to roomtemperature and stir overnight before quenching by the addition of EtOAcand water. The aqueous layer was extracted with EtOAc (3×300 mL). Thecombined organic layers were dried (MgSO₄), filtered, concentrated invacuo and the resulting residue purified by flash column chromatographyusing a gradient of 0 to 4% 2M NH₃ in EtOH/DCM to provide the desired109 (5.36 g, 76%). ¹H NMR (300 MHz, CDCl₃): δ 7.26-7.29 (m, 2H),7.12-7.16 (m, 2H), 3.58-3.64 (m, 1H), 3.47-3.53 (m, 1H), 3.00 (m, 1H),2.64 (s, 2H), 2.43-2.48 (m, 1H), 2.15-2.32 (m, 7H), 1.99-2.09 (m, 1H),1.79-1.92 (m, 1H), 1.36-1.69 (m, 4H), 1.16-1.29 (m, 1H). ¹³C NMR (75MHz, CDCl₃): δ 147.9, 131.0, 127.9, 127.4, 69.1, 67.6, 59.7, 56.2, 46.7,37.0, 31.8, 31.5, 27.2, 24.3, 15.9.

Alcohol 109 could be converted to the desired 114 using the procedureoutlined for the conversion of 36 to 37 in Example 22. Mesylateformation: 109 (7.25 g, 25 mmol), iPr₂NEt (10.90 mL, 63.0 mmol), MsCl(2.11 mL, 27 mmol), DCM (100 mL). After purification by flash columnchromatography using a gradient of 0 to 4% 2M NH₃ in EtOH/DCM thedesired mesylate was provided (8.42 g, 90%). Mesylate displacement:mesylate (8.42 g, 22.60 mmol), Cs₂CO₃ (18.40 g, 56.5 mmol), phenol (4.03g, 24.9 mmol), DMF (100 mL). Purification by silica gel columnchromatography using a gradient of 0 to 5% ethyl acetate/hexane providedthe desired 114. The enantiomeric excess could be determined via achiral AD column (100% MeOH) and was found to be 98%. LRMS: M+438. ¹HNMR (300 MHz, CDCl₃): δ 7.56 (2H, d, J=8.6 Hz), 7.23 (2H, d, J=8.5 Hz),7.10 (2H, d, J=7.7 Hz), 6.91 (2H, d, J=9.0 Hz), 4.74 (2H, m), 2.68-1.49(17H, m).

Example 76 Synthesis of(S)-1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-piperidine

115 was prepared from ethyl (S)-nipecotate D-tartrate and1-(4-chloro-phenyl)-cyclobutanecarboxylic acid, using the procedureoutlined in Example 75 for the synthesis of 114.

Preparation of 111: (S)-ethyl nipecotate L-tartrate (11.96 g, 38.8mmol), 1-(4-chlorophenyl)-1-cyclobutane carboxylic acid (12.23 g, 58mmol), PyBroP (27.16 g, 58.0 mmol), iPr₂NEt (34 mL, 194 mmol), DCM (170mL). Purification by flash column chromatography using 35% ethylacetate/hexane provided the desired amide 111 (7.73 g, 56%).

Preparation of 112: 111 (7.73 g, 22 mmol), LiAlH₄ (2.51 g, 66 mmol), THF(75 mL). Purification by flash column chromatography using a gradient of1 to 4% 2M NH₃ in EtOH/DCM provided the desired 112 (4.78 g, 74%).

Preparation of 115. Mesylate formation: amino alcohol (4.78 g, 16 mmol),iPr₂NEt (7.10 mL, 40 mmol), MsCl (1.40 mL, 18 mmol), DCM (66 mL). Afterpurification by flash column chromatography using a gradient of 1-4% 2MNH₃ in EtOH/DCM the desired mesylate was provided (4.74 g, 80%).Mesylate displacement: mesylate (4.74 g, 13.0 mmol), Cs₂CO₃ (10.6 g, 33mmol), phenol (2.27 g, 14 mmol), DMF (60 mL). The compound was purifiedusing silica gel chromatography using a gradient of 0 to 5% ethylacetate/hexane provided the desired 115. The enantiomeric excess couldbe determined via a chiral AD column (100% MeOH) and was found to be94%. LRMS: M+438. ¹H NMR (300 MHz, CDCl₃): δ 7.56 (2H, d, J=8.6 Hz),7.23 (2H, d, J=8.5 Hz), 7.10 (2H, d, J=7.7 Hz), 6.91 (2H, d, J=9.0 Hz),4.74 (2H, m), 2.68-1.49 (17H, m). ¹³C (partial, 100 MHz, CDCl₃): δ161.2, 144.4, 128.0, 127.8, 17.1, 114.6, 70.9, 69.0, 58.6, 36.2, 32.0,31.8, 26.9, 24.9, 16.3.

Example 77 Synthesis of 118 and 119

The two enantiomers of amide 38 were separated on a 2 cm AD chiralcolumn using 85% hexane (with 0.2% diethylamine)/15% isopropyl alcoholwith a flow rate of 6 mL/min. 116 retention time approx. 31 minutes.LRMS calculated for C₂₅H₂₇ClF₃NO₂ 465.17, found 465.55. 117 retentiontime approx. 41 minutes. LRMS calculated for C₂₅H₂₇ClF₃NO₂ 465.17, found465.68.

116 was reduced as per the procedure outlined for the reduction of amide38 to amine 39 in Example 22: 116 (39 mg, 0.0839 mmol), Red-Al (0.088mL, 0.294 mmol), toluene (0.5 mL). Purification by flash columnchromatography using 1% 2M NH₃ in EtOH/DCM provided 118 (18 mg, 47%).LRMS calculated for C₂₅H₂₉ClF₃NO_(451.19), found 451.28.

117 was reduced as per the procedure outlined for the reduction of amide38 to amine 39 in Example 22: 117 (38 mg, 0.0817 mmol), Red-Al (0.086mL, 0.286 mmol), toluene (0.5 mL). Purification by flash columnchromatography using 1% 2M NH₃ in EtOH/DCM provided 119 (15 mg, 41%).LRMS calculated for C₂₅H₂₉ClF₃NO_(451.19), found 451.85.

Example 78 Synthesis of(R)₁₋₂-(4-Chloro-phenyl)-2-methyl-propyl-3-(4-trifluoromethyl-phenoxymethyl)-piperidine)

2-(4-chlorophenyl)-2-methyl propionyl chloride was prepared as follows:To a flask containing 2-(4-chlorophenyl)-2-methyl propionic acid (15.0g, 75.6 mmol) was added thionyl chloride (approx. 6 mL). Dichloromethane(20 mL) was then added before adding an additional portion of thionylchloride (approx. 12 mL). The reaction was allowed to heat to 40° C. fortwo hours before cooling to room temperature and concentrating in vacuo(azeotroping with tetrahydrofuran). The resulting acid chloride was usedin the following acylation step without further purification orcharacterization.

To a solution of (R)-ethyl nipecotate L-tartrate (5.8 g, 18.9 mmol) insaturated aqueous NaHCO₃ (50 mL) was added a solution of2-(4-chlorophenyl)-2-methyl propionyl chloride (assume 100% yield fromabove preparation, 75.6 mmol) in ethyl acetate (50 m-L). The reactionwas allowed to stir at room temperature for two hours before furtherdiluting with ethyl acetate and saturated aqueous NaHCO₃. The layerswere separated and the aqueous layer further washed with ethyl acetate.The combined organic extracts were dried (MgSO₄), filtered, concentratedin vacuo and the resulting residue purified by flash columnchromatography using a gradient of 20 to 35% ethyl acetate/hexane toprovide the desired amide 120 (6.3 g, 99%).

The reduction of 120 to 121 could be accomplished as per the procedurefor the reduction of 108 to 109 outlined in Example 75 above: 120 (6.30g, 18.7 mmol), LiAlH₄ (2.13 g, 56.1 mmol), THF (85 mL). Purification byflash column chromatography using a gradient of 2 to 4% 2M NH₃ inEtOH/DCM provided the desired amide 121 (1.74 g, 82%). LRMS calculatedfor C₁₆H₂₄ClNO 281.15, found 281.96. ¹H NMR (300 MHz, CDCl₃): δ 7.32 (m,4H), 3.49-3.64 (m, 2H), 2.74 (m, 1H), 2.48-2.53 (m, 1H), 2.43 (s, 2H),2.14-2.31 (m, 3H), 1.65-1.73 (m, 2H), 1.53-1.64 (m, 1H), 1.41-1.50 (m,1H), 1.35 (s, 3H), 1.34 (s, 3H), 1.21 (m, 1H). ¹³C NMR (75 MHz, CDCl3):δ 147.0, 131.0, 127.8, 127.6, 71.3, 67.3, 59.8, 56.6, 39.1, 37.5, 27.0,26.8, 26.6, 24.6.

Alcohol 121 could be converted to the desired 122 using the procedureoutlined for the conversion of 36 to 37 in Example 22. Mesylateformation: 121 (4.48 g, 15.9 mmol), iPr₂NEt (6.94 mL, 39.8 mmol), MsCl(1.36 mL, 17.5 mmol), DCM (71 mL). After purification by flash columnchromatography using 2% 2M NH₃ in EtOH/DCM the desired mesylate wasprovided (5.71 g, 100%). Mesylate displacement: mesylate (5.71 g, 15.9mmol), Cs₂CO₃ (12.9 g, 39.6 mmol), α,α,α-trifluoro-p-cresol (2.57 g,15.9 mmol), DMF (81 mL). Purification by flash column chromatographyusing a gradient of 0.5 to 1% 2M NH₃ in EtOH/DCM provided the desired122 (2.67 g, 40%). LRMS calculated for C₂₃H₂₇ClF₃NO 425.17, found (M+)426.26. ¹H NMR (300 MHz, CDCl₃): δ 7.56 (d, J=8.7 Hz, 2H), 7.35 (d,J=8.7 Hz, 2H), 7.26 (d, J=8.7 Hz, 2H), 6.92 (d, J=8.7 Hz, 2H), 3.71-3.83(m, 2H), 2.50-2.55 (m, 1H), 2.35-2.45 (m, 3H), 2.13-2.20 (m, 1H),2.00-2.02 (m, 2H), 1.45-1.70 (m, 3H), 1.31 (s, 6H), 1.05-1.15 (m, 1H).¹³C NMR (75 MHz, CDCl₃): δ 161.5, 147.3, 131.2, 127.7, 126.8, 126.7,126.3, 122.5 (m), 114.3, 71.0, 70.6, 59.0, 56.6, 39.3, 36.1, 26.6, 26.5,24.7.

Example 79 Synthesis of[2-{3-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-cyclohexl}-2-(4-trifluoromethyl-phenoxy)-ethyl]-piperidine

Oxalyl chloride (18.5 mL) in 250 mL of CH₂Cl₂ was cooled down to −78°C., and DMSO (22.7 mL) was added slowly. The reaction mixture wasstirred for 10 minutes. The primary alcohol (30.0 g) in 300 mL of CH₂Cl₂was added dropwise to the cooled stirring reaction mixture. Aftercompletion of addition the reaction mixture was stirred for anadditional 15 minutes. At last, triethylamine (66.0 mL) was addedslowly. The reaction mixture was warmed to r.t. and stirred for 2 hours.The reaction mixture was washed with 500 mL of brine, 1.0 M NaHSO₄(2×100 mL), dried over anhydrous Na₂SO₄, and filtered. After removal ofthe solvent, the aldehyde was purified by a flash column chromatography(silica gel, Hexane/EtOAc, 8:2, yield, 95%).

Sodium hydride (4.12 g, 60% in mineral oil) in 100 mL of DMSO was heatedat 55° C. for 90 minutes and then cooled down to 0° C. A solution oftrimethylsulfonium iodide (21.62 g) in 100 mL of THF was added dropwiseand the resulting mixture was stirred for additional 15 minutes. Thealdehyde (10.0 g) in 100 mL of DMSO was then added. The reaction mixturewas stirred first at 0° C. for 15 minutes then at r.t. for 90 minutes.The mixture was quenched with 50 mL of water and extracted with hexane(3×100 mL). The combined organic layer was washed with brine, dried overanhydrous Na₂SO₄, filtered, and concentrated. The epoxide was used inthe next step without purification (yield: 94%).

The epoxide (1.0 g) was dissolved in 50 mL of piperidine in a sealedtube. The reaction mixture was heated at 70° C. overnight. Removal ofexcess piperidine gave the secondary alcohol (1.37 g). LRMS 312. Thecrude product was taken to the next step without further purification.

To the secondary alcohol (1.37 g) in 10 mL of CH₂Cl₂ was added 1.86 mLof N,N-Diiso-propylethylamine (2 eq.). The mixture was cooled down to 0°C., then methanesulfonyl chloride (0.619 mL, 1.5 eq.) was added. Thereaction mixture was stirred at r.t. for 2 hours and the solvent wasremoved. The crude residual was dissolved in 20 mL of CH₃CN and 3.66 gof potassium carbonate (5 eq.) and 1.72 g of α,α,α,trifluoro-p-cresol (2eq.) were added. The mixture was heated at 60° C. overnight. Thereaction mixture was quenched with 10% NaOH (5 mL) and extracted withEtOAc (3×10 mL). The combined organic layer was washed with brine anddried over Na₂SO₄. After filtration and removal of the solvent, theflash column chromatography (silica gel, Hexane/EtOAc, 4:1) gave thephenyl ether as a colorless oil (400 mg, LRMS 456).

To the phenyl ether (400 mg) in 1 mL of CH₂CL₂, 1.5 mL ofTrifluoroacetic acid was added at 0° C. The mixture was stirred at r.t.for 1 hour. The solvent was removed. 5 mL of 10% NaOH was added andaqueous solution was extracted with EtOAc (3×10 mL). The organic layerwas washed with brine and dried over Na₂SO₄. After removal of thesolvent, the residual was used in the next step. The crude productobtained from previous step (313 mg), potassium carbonate (608 mg, 5eq.) and 1-(1-Bromomethyl-cyclobutyl)-4-chloro-benzene (685 mg, 3 eq.),were dissolved in 2.0 mL of CH₃CN. The reaction mixture was stirred at70° C. overnight. The reaction mixture was quenched with 10% NaOH (5 mL)and extracted with EtOAc (3×10 mL). The combined organic layer waswashed with brine and dried over Na₂SO₄. After filtration and removal ofthe solvent, the preparative TLC (silica gel, hexane/EtOAc 7:3) gave1-[2-{3-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-cyclohexyl}-2-(4-trifluoromethyl-phenoxy)-ethyl]-piperidineas a colorless oil (LRMS 535).

Example 80 Synthesis of (S)-piperidine-1,3-dicarboxylic acid 1 benzylester 3-ethyl Ester

A 250 mL round-bottom flask was charged with K₂CO₃ (13.5 g, 97.6 mmol),piperidine (10 g, 32.5 mmol), and a 1:1 mixture of THF/H₂O (100 mL). A50 mL addition funnel was placed on the flask and charged with CbzCl(6.67 g, 39.0 mmol). The flask was cooled to 0° C. and then CbzCl wasadded dropwise over 5 minutes. The reaction mixture was warmed to 20° C.and stirred for 12 h. The reaction mixture was extracted with EtOAc (250mL) and the organic layer was washed with saturated NaCl (250 mL), dried(MgSO₄), filtered and concentrated in vacuo. The crude material (9.5 g,100% yield) was carried on without further purification.

Example 81 Synthesis ofS-3-Methanesulfonyloxymethyl-piperidine-1-carboxylic Acid Benzyl Ester

A 250 mL round-bottom flask was charged with alcohol (4.17 g, 16.7mmol), DCM (100 mL) and diisopropylethylamine (7.3 mL (42 mmol). Theflask was cooled to 0° C. and methanesulfonyl chloride (1.55 mL, 20mmol) was added dropwise. The reaction was warmed to 20° C. and stirredfor 12 h. The reaction mixture was diluted with DCM (150 mL). Theorganic layer was washed with 5% HCl (250 mL), saturated NaCl (250 mL),dried (MgSO₄), filtered and concentrated in vacuo. The crude materialwas purified by flash chromatography (silica gel, hexanes/EtOAc 2:1) togive pure product (5.46 g, 100% yield).

Example 82 Synthesis ofS-3-(4-Trifluoromethyl-phenoxymethyl)-piperidine-1-carboxylic AcidBenzyl Ester

A 250 mL round-bottom flask was charged with mesylate (4.17 g, 12.7mmol), α,α,α-trifluoromethyl-p-cresol (2.27 g, 14.0 mmol), MeCN (100 mL)and Cs₂CO₃ (10.4 g, 31.8 mmol). The reaction mixture was heated toreflux for 12 h. The reaction mixture was cooled and diluted with EtOAc(250 mL). The organic layer was washed with H₂O (250 mL), saturated NaCl(250 mL), dried (Na₂SO₄), filtered and concentrated in vacuo. The crudematerial was purified by flash chromatography (silica gel, hexanes/EtOAc3:1) to give pure product (3.97 g, 79% yield).

Example 83 Synthesis of S-3-(4-Trifluoromethyl-phenoxymethyl)-piperidine

A 100 mL round-bottom flask was charged with Cbz-protected amine (4.0 g,10 mmol) and methanol (20 mL). The flask was blanketed under argon and10% w/w palladium on carbon (1.08 g, 1 mmol) was added. The flask wasplaced under hydrogen gas (1 atmosphere) and stirred for 3 h. Thereaction mixture was filtered through celite and concentrated in vacuoto give crude product (2.91 g, 79% yield) that was used withoutpurification.

Example 84 Synthesis ofS-1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone

A 50 mL pear-bottom flask was charged with α-chloroketone (700 mg, 2.9mmol), acetone (10 mL) and sodium iodide (432 mg, 2.9 mmol). Thereaction mixture was heated to reflux for 10 minutes and then a solutionof amine (622 mg, 2.4 mmol) in acetone (15 mL) was added followed byCs₂CO₃ (1.6 g, 4.8 mmol). The reaction mixture was heated to reflux for6 h and then diluted with EtOAc (50 mL). The organic layer was washedwith saturated NaCl (50 mL), dried (Na₂SO₄), filtered and concentratedin vacuo. The crude material was purified by flash chromatography(silica gel, hexanes/EtOAc 85:15 w/5% 2.0 M NH₃ in EtOH) to give 123(622 mg, 56% yield).

Example 85 Synthesis of1R-1-[1-(4-Chloro-phenyl):cyclobutyl]-2-[(3S)-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanoland1S-1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[(3S)-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A 100 mL round-bottom flask was charged with 123 (622 mg, 1.33 mmol),methanol (10 mL) and sodium borohydride (56 mg, 1.47 mmol). The reactionmixture was stirred for 2 h and then quenched with H₂O (10 mL) andextracted with EtOAc (50 mL). The organic layer was washed with 5% HCl(50 mL), saturated NaCl (50 mL), dried (Na₂SO₄), filtered andconcentrated in vacuo. The crude material was purified by flashchromatography (hexanes/EtOAc 1:1 w/5% 2.0 M NH₃ in EtOH) to give thealcohol as a 1:1 mixture of diastereomers (534 mg). The diastereomerswere separated by first recrystallization from hot methanol to give 124(140 mg) followed by preparatory HPLC (chiral AD column,hexanes/ethanol/diethylamine 95:5:0.1) to give additional 124 (70 mg,RT=33 min) and 125 (212 mg, RT=39 min).

Example 86 Synthesis of1R-1-[1](4-Chloro-phenyl)-cyclobutyl]-2-[(3R)-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanoland1S-1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[(3R)-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

Synthesis ofS-1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone

A solution of amine 74 (7.6 g) and potassium carbonate (8.40 g) inacetone (50 mL) was stirred at RT for 30 min. The α-chloroketone (5 g)dissolved in acetone (50 mL) was stirred at RT in a separate reactionvessel. After 5 min. of stirring, the α-chloroketone solution was addedto the reaction mixture containing the amine and potassium carbonate.After completion of addition the reaction mixture was heated to 50 C.After 18 h the reaction mixture was poured into water (400 mL) and theaqueous layer was extracted with EtOAc. Combined organics were dried(Na₂SO₄), filtered and concentrated in vacuo. The crude material waspurified by flash chromatography (silica gel, hexanes/EtOAc 80:16 w/4%2.0 M NH₃ in EtOH) to give 128 (8 g).

Synthesis of1R-1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[(3R)-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanoland1S-1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[(3R)-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A 100 mL round-bottom flask was charged with 128 (4.65 g, 9.98 mmol),methanol (69 mL) and sodium borohydride (566 mg, 14.98 mmol). Progressof the reaction was monitored by TLC. Upon completion, the reactionmixture was concentrated to yield a yellow residue. This residue wastaken up in EtOAc and then diluted with water. The aqueous layer wasextracted with EtOAc (3×50 mL). The combined organic layers were dried(Na₂SO₄), filtered and concentrated in vacuo. The crude material waspurified by flash chromatography (80:16: 4 hexane: EtOAc: 2.0 M NH₃ inEtOH) to give the alcohol as a 1:1 mixture of diastereomers (2.5 g). Thediastereomers were separated by first recrystallization from methanol togive isomer 126-followed by preparatory HPLC (chiral AD column,hexanes/ethanolidiethylamine 95:5:0.1) to give additional pure isomer126 (RT=28 min) and pure isomer 127 (RT=45 min).

Example 87 Synthesis ofS-1-[1-(4-Chloro-phenyl)-cyclobutanecarbonyl]-piperidine-3-carboxylicAcid Ethyl Ester

The acid chloride was prepared from the corresponding acid (8.5 g, 40/7mmol) by treatment with oxalyl chloride (35.5 mL, 407 mmol) and DMF (1drop) in a 50 mL round-bottom flask at 20° C. for 3 h. The acid chloridewas concentrated in vacuo and used immediately. A 250 mL round-bottomflask was charged with amine tartrate (2.5 g, 8.16 mmol), EtOAc (50 mL)and saturated NaHCO₃ (50 mL). Acid chloride (40.7 mmol) was added whilestirring vigorously. The reaction was stirred for 1 h and then theorganic layer was separated, dried (Na₂SO₄), filtered and concentratedin vacuo. The crude material was purified by flash chromatography(silica gel, hexanes/EtOAc 3:1) to give pure product (2.35 g, 82%yield).

Example 88 Synthesis ofS-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-methanol

A 100 mL round-bottom flask was charged with ester (2.35 g, 6.7 mmol)and THF (30 mL). The reaction flask was cooled to 0° C. and a 1.0 Msolution of LAH in THF (20.2 mL, 20.2 mmol) was added dropwise. Thereaction mixture was allowed to warm to 20° C. and stirred for 12 h. Thereaction mixture was quenched slowly with 10% HCl (1 mL). The pH wasadjusted to 8 with 10% NaOH and extracted with EtOAc (2×50 mL). Theorganic layer was washed with saturated NaCl (50 mL), dried (Na₂SO₄),filtered and concentrated in vacuo. The crude material was purified byflash chromatography (silica gel, DCM w/5% 2.0 M NH₃ in EtOH) to givepure product (1.9 g, 88% yield).

Example 89 Synthesis ofS-1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-benzyloxymethyl)-piperidine

A 100 mL round-bottom flask was charged with NaH (237 mg, 7.76 mmol) andTHF (25 mL). The reaction flask was cooled to 0° C. and alcohol wasadded (1.9 g, 6.5 mmol). The reaction mixture was stirred for 15 minutesand then benzylbromide (1.86 g, 7.76 mmol) was added. The reaction washeated to 60° C. and stirred for 12 h. The reaction mixture was cooledto 20° C., quenched with water (25 mL) and extracted with EtOAc (50 mL).The organic layer was washed with saturated NaCl (50 mL), dried(Na₂SO₄), filtered and concentrated in vacuo. The crude material waspurified by flash chromatography (silica gel, hexanes/EtOAc 3:1) to give129 (2.3 g, 78% yield). ¹H-NMR (CDCl₃) (300 MHz) δ 7.65 (d, 2H), 7.42(d, 2H), 7.24 (d, 2H), 7.15 (d, 2H), 5.51 (s, 2H), 3.24 (m, 2H), 2.63(s, 2H), 2.50 (d, 1H), 2.40-2.18 (m, 5H), 2.07 (m, 2H), 1.96 (m, 3H),1.58 (m, 1H), 1.43 (m, 2H), 0.98 (m, 1H). ¹³C-NMR (CDCl₃) (300 MHz) δ148.7, 143.2, 131.0, 127.9, 127.8, 127.6, 125.6, 125.5, 74.1, 72.3,69.0, 59.6, 56.5, 47.4, 36.9, 31.8, 27.3, 25.1, 16.3. MS (APCI) m/z451.8 [MH]⁺.

Example 90 Synthesis ofR-1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-benzyloxymethyl)-piperidine

R-1-[1-(4-Chloro-phenyl)-cyclobutanecarbonyl]-piperidine-3-carboxylicAcid Ethyl Ester

A 250 mL round-bottom flask was charged with amine tartrate (3.44 g,11.2 mmol), EtOAc (50 mL) and saturated NaHCO₃ (50 mL). Acid chloride(56.0 mmol) was added while stirring vigorously. The reaction wasstirred for 1 h and then the organic layer was separated, dried(Na₂SO₄), filtered and concentrated in vacuo. The crude material waspurified by flash chromatography (silica gel, hexanes/EtOAc 3:1) to givepure product (3.8 g, 97% yield)

R-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-methanol

A 100 mL round-bottom flask was charged with ester (3.80 g, 10.9 mmol)and THF (25 mL). The reaction flask was cooled to 0° C. and a 1.0 Msolution of LAH in THF (32.6 mL, 32.6 mmol) was added dropwise. Thereaction mixture was allowed to warm to 20° C. and stirred for 12 h. Thereaction mixture was quenched slowly with 10% HCl (1 mL). The pH wasadjusted to 8 with 10% NaOH and extracted with EtOAc (2×50 mL). Theorganic layer was washed with saturated NaCl (50 mL), dried (Na₂SO₄),filtered and concentrated in vacuo. The crude material was purified byflash chromatography (silica gel, DCM w/5% 2.0 M NH₃ in EtOH) to givepure product (2.8 g, 88% yield).

R-1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-benzyloxymethyl)-piperidine

A 100 mL round-bottom flask was charged with NaH (348 mg, 11.4 mmol) andTHF (35 mL). The reaction flask was cooled to 0° C. and alcohol (2.8 g,9.5 mmol) was added. The reaction mixture was stirred for 15 minutes andthen benzylbromide (2.73 g, 11.4 mmol) was added. The reaction washeated to 60° C. and stirred for 12 h. The reaction mixture was cooledto 20° C., quenched with water (25 mL) and extracted with EtOAc (50 mL).The organic layer was washed with saturated NaCl (50 mL), dried(Na₂SO₄), filtered and concentrated in vacuo. The crude material waspurified by flash chromatography (silica gel, hexanes/EtOAc 3:1) to give130 (3.1 g, 73% yield). ¹H-NMR (CDCl₃) (300 MHz) δ 7.65 (d, 2H), 7.42(d, 2H), 7.24 (d, 2H), 7.15 (d, 2H), 5.51 (s, 2H), 3.24 (m, 2H), 2.63(s, 2H), 2.50 (d, 1H), 2.40-2.18 (m, 5H), 2.07 (m, 2H), 1.96 (m, 31),1.58 (m, 1H), 1.43 (m, 2H), 0.98 (m, 1H). ¹³C-NMR (CDCl₃) (300 MHz) δ148.7, 143.2, 131.0, 127.9, 127.8, 127.6, 125.6, 125.5, 74.1, 72.3,69.0, 59.6, 56.5, 47.4, 36.9, 31.8, 27.3, 25.1, 16.3. MS (APCI) m/z451.8 [MH]⁺.

Example 91 Synthesis ofcis-1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-2-yl]-methanol,as a Racemate and as Single Enantiomers (Absolute StereochemistryRandomly Assigned to all Single Enantiomers)

3-Hydroxymethyl-pyridine-2-carboxylic acid isopropyl ester (1.0 g, 5.1mmol, prepared as in Ornstein, et al., J. Med. Chem. 1989, 32, 827) wasconverted to its t-butyldimethylsilyl ether under standard conditions(5.6 mmol Me₂Si(t-Bu)Cl, 11.2 mmol imidazole, 25 mL DMF, roomtemperature, overnight). Dilution with 50 mL of water, addition of 100mL ether, and extractive workup gave, after concentration of the organiclayers in vacuo and chromatography on silica gel, the desired compound,3-(tert-Butyl-dimethyl-silanyloxymethyl)-pyridine-2-carboxylic acidisopropyl ester (1.15 g, 73%). MS 310 (M+1); ¹H NMR (300 MHz, CDCl₃): δ8.59 (br. d, J=4.6 Hz, 1H), 8.17 (br.d, J=8.0 Hz, 1H), 7.41 (dd, J=8.0,4.6 Hz, 1H), 5.22 (m, 1H), 5.02 (s, 2H), 1.39 (d, J=6.4 Hz, 6H), 0.91(s, 9H), 0.03 (s, 6H).

3-(tert-Butyldimethyl-silanyloxymethyl)-pyridine-2-carboxylic acidisopropyl ester (800 mg, 2.6 mmol) was dissolved in 6 mL of methanol ina pressure hydrogenation vessel to which 100 mg of PtO₂ was added (Rh onalumina may also be used). The vessel was shaken under 50 psi ofhydrogen for 5 hrs. The suspension was filtered through Celite and thesolution was concentrated in vacuo to provide the desired compound,cis-3-(tert-butyl-dimethyl-silanyloxymethyl)-piperidine-2-carboxylicacid isopropyl ester. MS 316 (M+1). Without further purification theproduct was acylated under standard conditions with1-(4-chloro-phenyl)-cyclobutanecarbonyl chloride (itself prepared fromthe carboxylic acid using excess thionyl chloride at reflux for 1 hr.followed by concentration in vacuo). Water/ether extractive workup gave,after concentration of the organic layers in vacuo and chromatography onsilica gel, the desired compound,cis-3-(tert-Butyl-dimethyl-silanyloxymethyl)-1-[1-(4-chloro-phenyl)-cyclobutanecarbonyl]-piperidine-2-carboxylicacid isopropyl ester (720 mg, 55% for two steps) MS 508 (M+1). The cisstereochemistry was assigned based upon a ¹H NMR coupling constant of 4Hz for the proton at C-2 of the piperidine ring.

Cis-3-(tert-Butyl-dimethyl-silanyloxymethyl)-1-[1-(4-chloro-phenyl)-cyclobutanecarbonyl]-piperidine-2-carboxylicacid isopropyl ester (460 mg, 0.9 mmol) was dissolved in 10 mL of THF.Lithium aluminum hydride (171 mg, 4.5 mmol) was added slowly and thesuspension was brought to reflux for 1 hr. The suspension was cooled to0° C. and cold 0.5 M NaOH (0.75 mL) was added dropwise. The slurry wasvigorously stirred at room temperature for 30 minutes, filtered throughCelite, and the solution was concentrated in vacuo to provide, afterchromatography on silica gel, the desilylated and reduced compound,cis-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-hydroxymethyl-piperidin-2-yl}-methanol(176 mg, 60%) MS 324 (M+1).

Cis-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-hydroxymethyl-piperidin-2-yl}-methanol(95 mg, 0.29 mmol) was dissolved in 5 mL of THF. KOtBu (0.58 mmol, 66mg) and 1-fluoro-4-trifluoromethyl-benzene (0.29 mmol, 48 mg) was addedand the solution was brought to reflux for 4 hours. Water/etherextractive workup gave, after concentration of the organic layers invacuo and chromatography on silica gel, the desired mono-arylatedcompound,cis-[1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-2-yl]-methanol(82) (39 mg, 28%). The enantiomeric mixture of compounds was separatedby preparative HPLC using a Chiralpak OD™ column from ChiralTechnologies, Inc., eluting with an 85:15 mixture of hexane andisopropyl alcohol containing ca 0.2% diethylamine. The products (131 and132) were isolated and converted individually into their HCl salts byexposure to a solution of HCl in ether. Data for HCl salt form: MS 468(M+1). ¹H NMR (300 MHz, CDCl₃): δ 9.23 (br. s, 1H, NH of protonatedtertiary amine), 7.56 (d, J=7.4 Hz, 2H), 7.39-7.49 (m, 4H), 6.85 (d,J=7.4 Hz, 2H), 3.6-3.95 (br. m, 7H), 1.45-3.25 (br. overlappingmultiplets, total 14H). ¹³C NMR (CDCl₃, 75 MHz): δ 160.4, 143.0, 133.5,129.7, 128.2, 127.3 (CF₃), 124.2, 114.8, 114.6, 68.3, 65.0, 63.4, 56.0,50.6, 45.1, 33.9, 32.9, 30.9, 21.5, 19.1, 16.1.

To confirm the identity of the final products as the enantiomers ofcis-[1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-2-yl]-methanolrather than the regioisomers[1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-2-(4-trifluoromethyl-phenoxymethyl)-piperidin-3-yl]-methanol,the Scheme 1 synthesis was modified as shown in Scheme 2. Experimentalprocedures employed are standard and generally follow proceduresoutlined for Scheme 1 with the exception that the hydroxyl “protectinggroup” strategies are altered in a straightforward fashion. Thismodified route uses an ethoxyethyl group rather than atert-butyl-dimethyl-silanyl group for blocking the hydroxyl function ofthe starting material 3-hydroxymethyl-pyridine-2-carboxylic acidisopropyl ester. The ethoxyethyl group, unlike thetert-butyl-dimethyl-silanyl group, is stable to the lithium aluminumhydride reduction. By employing a triisopropyl-silanyl group for thefunctionalization of the hydroxymethyl group at the 2 position of thepiperidine system, the alcohol functions remain differentially blocked.Thus it was unambiguously determined that the aryl ether present in thefinal product is off the 3 position rather than the two position of thepiperidine ring system. This modified synthesis ofcis-1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-2-yl]-methanol(82 as the racemate, 131 and 132 as single enantiomers) is shown inScheme 2.

3-Hydroxymethyl-pyridine-2-carboxylic acid isopropyl ester (5.2 g, 26.6mmol) prepared as in Ornstein, et al., J. Med. Chem. 1989, 32, 827) wasconverted to ethoxyethyl ether under standard conditions (excess ethylvinyl ether, ca. 10 mL; 75 mL dry methylene chloride, catalyticpyridinium p-toluene sulfonate). Dilution with 100 mL of water, additionof 250 mL ether, and extractive workup gave, after concentration of theorganic layers in vacuo and chromatography on silica gel, the desiredcompound 3-(1-Ethoxy-ethoxymethyl)-pyridine-2-carboxylic acid isopropylester, 6.13 g (86%). Data: MS 268 (M+1); ¹H NMR (300 MHz, CDCl₃): δ 8.63(br. d, J=4.6 Hz, 1H), 8.09 (br. d, J=7.9 Hz, 1H), 7.43 (dd, J=7.9, 4.6Hz), 5.17 (m, 1H), 5.02 and 4.94 (AB d, J=14.9 Hz, 2H), 4.81 (q, J=7.0Hz, 1H), 3.50-3.75 (m, 2H), 1.40-1.45 (overlapping doublets, 9H), 1.21(t, J=7.0 Hz, 3H); ¹³C NMR (CDCl₃, 75 MHz): δ 165.7, 147.9, 146.3,136.8, 136.1, 126.0, 100.1, 69.6, 63.7, 61.4, 21.9, 20.1, 15.4.

Hydrogenation of compound3-(1-ethoxy-ethoxymethyl)-pyridine-2-carboxylic acid isopropyl ester,following the method used for the corresponding silyl ether as in scheme1, gave the desired product3-(1-ethoxy-ethoxymethyl)-piperidine-2-carboxylic acid isopropyl ester.The starting pyridine compound (5.1 g, 19.1 mmol) was dissolved in 60 mLof methanol in a pressure hydrogenation vessel to which 2 g of PtO₂ wasadded. The vessel was shaken under 60 psi of hydrogen for 5 days. Thesuspension was filtered through Celite and the solution was concentratedin vacuo to provide the desired compound,3-(1-ethoxy-ethoxymethyl)-piperidine-2-carboxylic acid isopropyl ester(5.1 g, ca. 100%). Data: MS 274 (M+1); ¹H NMR (300 MHz, CDCl₃): δ 5.50(br. s, 1H, NH), 5.07 (m, 1H, CH of ester), 4.60-4.64 (overlapping q's,J=7.0 Hz, 1H), 3.41-3.73 (m, 5H), 3.10 (m, 1H), 2.69 (m, 1H), 2.30 (m,1H), 1.80-1.95 (m, 2H), 1.41-1.63 (m, 3H), 1.19-1.30 (m, 12H); ¹³C NMR(CDCl₃, 75 MHz): δ 172.4, 172.3, 100.0, 99.9, 68.1, 68.0, 64.4, 63.7,61.1, 61.0, 60.4, 46.0, 45.9, 36.81, 36.76, 26.3, 26.1, 22.14, 22.07,22.0, 21.9, 19.92, 19.85, 15.45, 15.39.

Acylation and LAH reduction, were accomplished following the sameprocedures used in the scheme 1 route, affording the desired products in88% (15 mmol scale of the amine) and 73% yields (13.5 mmol scale of theamide), respectively. Mass spectral data for these intermediates are asfollows:1-[1-(4-chloro-phenyl)-cyclobutanecarbonyl]-3-(1-ethoxy-ethoxymethyl)-piperidine-2-carboxylicacid isopropyl ester MS 466 (M+1),[1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(1-ethoxy-ethoxymethyl)-piperidin-2-yl]-methanolMS 396 (M+1). Protection of the alcohol function as thetriisopropylsilyl ester followed the procedure for thet-butyldimethylsilyl ester protection in scheme 1 (92 yield, 12.2 mmolscale of the alcohol,1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-3-(1-ethoxy-ethoxymethyl)-2-triisopropylsilanyloxymethyl-piperidineMS 552 (M+1). Hydrolysis of the ethoxyethyl function (11.1 mmol startingether, 50 mL MeOH, 5 mL TFA, 30 min, concentrated in vacuo) gave thefree alcoholChloro-phenyl)-cyclobutylmethyl]-2-triisopropylsilanyloxymethyl-piperidin-3-yl}-methanolin 88% yield after chromatographic purification on silica gel. Data: MS480 (M+1). This alcohol (960 mg, 2.0 mmol) was dissolved in 15 mL ofTHF. KOtBu (4.1 mmol) and 1-Fluoro-4-trifluoromethyl-benzene (5.1 mmol)was added and the solution was brought to reflux for 4 hours.Water/ether extractive workup gave, after concentration of the organiclayers in vacuo and chromatography on silica gel, the desired arylatedcompound1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-2-triisopropylsilanyloxymethyl-piperidine(968 mg, 78%); Data: MS 624 (M+1). Finally, this compound (746 mg, 1.19mmol) was dissolved in 5 mL of tetrahydrofuran and 1.5 mL of a 1Msolution of tetrabutylammonium fluoride in THF was added dropwise viasyringe. The solution was stirred overnight at ambient temperature.Water/ether extractive workup gave, after concentration of the organiclayers in vacuo and chromatography on silica gel, the desired compound82. The final products obtained via the route in Scheme 2 (131 and 132,obtained after HPLC purification as described above, 82 as the racemate)were identical in all respects to products obtained via the route inScheme 1.

Example 92 Synthesis of(R)-1-[1-(4-Chloro-phenyl)-cyclobutanecarbonyl]-piperidine-3-carboxylicAcid Ethyl Ester

Ethyl (R)-nipecotate L-tartrate (15.0 g, 48.8 mmol) was added to astirred mixture of dichloromethane and saturated sodium bicarbonatesolution (100 mL each) at 0° C. After 10 min1-(4-chloro-phenyl)-cyclobutanecarbonyl chloride (11.13 g, 48.8 mmol)was slowly added at 0° C. The reaction was then allowed to stir for 2 hat room temperature. The organic layer was separated and the aqueouslayer was washed once with dichloromethane (100 mL). The organicmaterial was combined and washed once with water (100 mL), dried overanhydrous sodium sulfate, filtered and concentrated by rotaryevaporation. The residue was purified by chromatography on silica gel,eluting with hexane/ethyl acetate (2:1) to give 9.2 g of the amidesteras a thick clear gum; C₁₉H₂₄ClNO₃, LRMS (m/z)=350 (MH+).

Example 93 Synthesis of(R)-[1-(4-Chloro-phenyl)-cyclobutyl]-(3-hydroxymethyl-piperidin-1-yl)-methanone

To a solution of the amidester (6.0 g, 17.2 mmol) in absolute ethanol(100 mL) at 0° C. was slowly added sodium borohydride (0.65 g, 17.2mmol). The reaction was then allowed to warm to room temperature andstirred overnight. The reaction was quenched by slow addition of water,and then most of the ethanol was removed by rotary evaporation. Theresidue was partitioned between dichloromethane and water (100 mL each),and the aqueous layer was extracted once with dichloromethane (100 mL).The organic material was combined and dried over anhydrous sodiumsulfate, filtered and concentrated by rotary evaporation. The residuewas purified by chromatography on silica gel, eluting withdichloromethane/methanol (98:2) to give 4.9 g of the alcohol as a thickclear gum; C₁₇H₂₂ClNO₂, LRMS (m/z)=308 (MH+).

Example 94 Synthesis of Methanesulfonic Acid(R)-1-[1-(4-chloro-phenyl)-cyclobutanecarbonyl]-piperidin-3-ylmethylEster

The alcohol derivative (4.5 g, 14.7 mmol)) was dissolved indichloromethane (100 mL) and cooled to 0° C. To this solution was addedtriethylamine (5.0 mL) followed by dropwise addition of methanesulfonylchloride (1.68 g, 14.7 mmol) at 0° C. The reaction was stirred at 0° C.for 1 h and then at room temperature for 4 h. The reaction mixture waswashed successively with water, 1 N HCl, water, sat. sodium bicarbonatesolution, and water (100 mL each). The organic layer was dried overanhydrous sodium sulfate, filtered and concentrated by rotaryevaporation. The residue gave 5.0 g of the mesylate as a thick yellowgum which was used in the following step without further purification;C₁₈H₂₄ClNO₄S, LRMS (m/z)=386 (MH+).

Example 95 Synthesis of(R)-[1-(4-Chloro-phenyl)-cyclobutyl]-[3-(4-fluoro-phenoxymethyl)-piperidin-1-yl]methanone

To a solution of the mesylate (4.0 g, 10.4 mmol) in acetonitrile (100mL) was added 4-fluorophenol (1.17 g, 10.4 mmol) and cesium carbonate(3.40 g, 10.4 mmol). The reaction was stirred and refluxed for 20 h.After cooling to room temperature, the reaction mixture was filtered andmost of the solvent was removed by rotary evaporation. The residue waspartitioned between dichloromethane and water, and the organic layer waswashed with sat. sodium carbonate (2×50 mL) and water. The organic layerwas dried over anhydrous sodium sulfate, filtered and concentrated byrotary evaporation. The residue was purified by chromatography on silicagel, eluting with dichloromethane/methanol (98:2) to give 3.2 g of theamide as a white solid; C₂₃H₂₅ClFNO₂, LRMS (m/z)=402 (MH+).

Example 96 Synthesis of(R)-1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(4-fluoro-phenoxymethyl)-piperidine

To a suspension of lithium aluminum hydride (0.19 g, 5.0 mmol) inanhydrous tetrahydrofuran (50 mL) at 0° C. was added the amide (2.0 g,4.99 mmol). The reaction mixture was then stirred at 0° C. for 4 h. Thereaction mixture was then quenched at 0° C. with slow addition of waterand 1 N NaOH. The residue was extracted well with ethyl acetate (4×100mL), and the combined organic portions were dried over anhydrousmagnesium sulfate, filtered and concentrated by rotary evaporation. Theorganic residue was purified by flash chromatographey on silica gel,eluting with dichloromethane/2.0 M ammonia in ethyl alcohol (98:2) togive 1.35 g of 133 as a pale yellow oil; C₂₃H₂₇ClFNO, LRMS (m/z)=388(MH+). Enantiomeric excess was determined via Chiral HPLC using aChiralpak®AD Column (Chiral Technologies, Inc.; 10 μm, 4.6 mm I.D.×250mm) eluting with methanol/water/diethylamine (95:5:0.1) with a flow rateof 1.0 ml/min. and was found to be >98% (retention time of 133:9.51min.).

Example 97 Synthesis of(R)-[1-(4-Chloro-phenyl)-cyclobutyl]-[3-(4-fluoro-phenoxymethyl)-piperidin-1-yl]-methanoneand(S)-[1-(4-Chloro-phenyl)-cyclobutyl]-[3-(4-fluoro-phenoxymethyl)-piperidin-1-yl]-methanone

Methanesulfonic Acid1-[1-(4-chloro-phenyl)-cyclobutanecarbonyl]-piperidin-3-ylmethyl Ester

To a solution of 3-piperidinemethanol (5.0 g, 43.4 mmol),1-(4-chlorophenyl)-1-cyclobutanecarboxylic acid (9.14 g, 43.4 mmol), anddiisopropylethylamine (11.22 g, 86.8 mmol) in dichloromethane (100 mL)at 0° C. was added PyBroP® (22.26 g, 47.8 mmol). The reaction wasstirred at 0° C. for 1 h and then at room temperature for 4 h. Thereaction mixture was washed successively with water, 1 N HCl, water,sat. sodium bicarbonate solution, and water (100 mL each). The organiclayer was dried over anhydrous sodium sulfate, filtered and concentratedby rotary evaporation. The residue was purified by chromatography onsilica gel, eluting with dichloromethane/methanol (96:4) to give 5.8 gof the hydroxyl amide as a thick gum.

The hydroxyl amide (5.0 g) was dissolved in dichloromethane (50 mL) andcooled to 0° C. To this solution was added pyridine (5.0 mL) followed bydropwise addition of methanesulfonyl chloride (2.05 g, 17.9 mmol). Thereaction was stirred at for 1 h and then at room temperature overnight.The reaction mixture was washed successively with water, 1 N HCl, water,sat. sodium bicarbonate solution, and water (100 mL each). The reactionmixture was washed successively with water, sat. sodium bicarbonatesolution, and water (100 mL each). The organic layer was dried overanhydrous sodium sulfate, filtered and concentrated by rotaryevaporation. The residue was purified by chromatography on silica gel,eluting with hexane/ethyl acetate (2:1) to give 4.45 g of the mesylateamide as a tan gum; C₁₈H₂₄ClNO₄S, LRMS (m/z)=386 (MH+).

[1-(4-Chloro-phenyl)-cyclobutyl]-[3-(4-fluoro-phenoxymethyl)-piperidin-1-yl]-methanone

To a solution of mesylate amide (1.0 g, 2.60 mmol) in acetonitrile (25mL) was added 4-fluorophenol (0.29 g, 2.60 mmol) and cesium carbonate(1.27 g, 3.90 mmol). The reaction was stirred and refluxed for 20 h.After cooling to room temperature, the reaction mixture was filtered andmost of the solvent was removed by rotary evaporation. The residue waspartitioned between dichloromethane and water, and the organic layer waswashed with sat. sodium carbonate (2×50 mL) and water. The organic layerwas dried over anhydrous sodium sulfate, filtered and concentrated byrotary evaporation. The residue was purified by chromatography on silicagel, eluting with dichloromethane/methanol (98:2) to give the etheramide (0.54 g, 52%) as a crystalline solid; C₂₃H₂₅ClFNO₂, LRMS (m/z)=402(MH+).

(R)-[1-(4-Chloro-phenyl)-cyclobutyl]-[3-(4-fluoro-phenoxymethyl)-piperidin-1-yl]-methanone,and(S)-[1-(4-Chloro-phenyl)-cyclobutyl]-[3-(4-fluoro-phenoxymethyl)-piperidin-1-yl]-methanone

Ether amide was separated via Chiral HPLC using a Chiralpak®AD Column(Chiral Technologies, Inc.; 21 mm I.D.×250 mm) eluting withhexane/2-propanol (85:15) to give enantiomers A and B.

Compounds 133 and 134 were separated via Chiral HPLC using aChiralpak®AD Column (Chiral Technologies, Inc.; 10 μm, 4.6 mm I.D.×250mm) eluting with methanol/water/diethylamine (95:5:0.1) with a flow rateof 1.0 ml/min. Retention time of 133:9.51 min. Retention time of134:10.8 min.

Example 98 Synthesis of Polymer Supported Methanesulfonic AcidPiperidin-3-ylmethyl Ester

To the Wang resin (12 g, 1.1 mmol/g) in a 250 mL peptide synthesisvessel was added 120 mL of 0.4 N CDI in anhydrous THF, and shaken atroom temperature for 17 hours. The resin was thoroughly washed withCH₂Cl₂ (3×100 mL) and THF (3×100 mL) to remove the excess CDI and thentreated with 120 mL of 0.4 N 3-piperidinemethanol in THF at roomtemperature for 17 hours. The resulting resin 1 was washed with DMF(3×100 mL), MeOH (4×100 mL), and CH₂Cl₂ (4×100 mL) and dried in vacuo.To the alcohol resin 1 was added methanesulfonyl chloride (5.11 ml, 66mmol) in 100 mL CH₂Cl₂ followed by 20 mL of pyridine, and the resultingslurry was shaken at room temperature for 17 hours. The resultingmesylate resin 2 was washed with DMF (3×100 mL), MeOH (4×100 mL), andCH₂Cl₂ (4×100 mL) and dried in vacuo.

Synthesis of 3-(4-Methoxy-phenoxymethyl)piperidine (1)

To mesylate resin 2 (1.0 g, 1.1 mmol) was added cesium carbonate (1.79g, 5.5 mmol) followed by 4-methoxyphenol (0.682 g, 5.5 mmol) in 10 mL ofDMF, and the mixture was shaken at 75° C. for 24 hours. The resultingresin 3 was extensively washed with DMF (3×10 mL), water (3×10 mL), MeOH(4×10 mL), and CH₂Cl₂ (4×10 mL). The resin was dried in vacuo andtreated with a solution of 50% TFA in CH₂Cl₂ at room temperature for 30min to release polymer-bound 3-(4-Methoxy-phenoxymethyl)piperidine.Removal of the volatiles under a stream of nitrogen followed by dryingin vacuo afforded 1 as a TFA salt, LRMS m/z 222. Piperidine derivatives2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 were prepared using the samegeneral procedure.

Synthesis of Combinatorial Library of Compounds of the Present Invention

Twelve piperidines (1 to 12) were dissolved in TMOF (4 mL), andrespectively dispensed into 96-well reaction block from column 1 tocolumn 12 at 0.5 ml/well (Scheme 1). Eight aldehydes (A to H) in TMOFwere respectively dispensed into eight rows, from row A to row H at 0.33ml/well (containing 0.11 mmol aldehyde). The mixtures were shaken at rtfor one hour, then (polystyrylmethyl)trimethylammonium cyanoborohydride(4.25 mmol/g) was dispensed into 96 wells at 0.1 g/well. The mixtureswere shaken at rt for 24 hours. Eight aldehydes were dispensed againinto the reaction block, and the shaking continued for another 24 hours.The reaction mixtures were filtered and the resins were washed with MeOH(3×0.5 ml/well). After conditioning SPE columns (SCX cation exchange,0.5 g of sorbent, 2.0 mequiv/g) with 5 mL of MeOH, the reaction contentswere loaded onto the column. The column was washed with 2×5 mL of MeOH,and eluted with 4 mL of 2.0 M ammonia in MeOH. The effluents werecollected into receiving tubes, concentrated and dried in vacuo toafford 96 final compounds, which were submitted to HPLC and mass spectraanalyses.

SCHEME 1 DAT Library

Example 99

The compounds prepared in the combinatorial library in Example 98 werescreened for their ability to inhibit the uptake of human monoamines.The ability of test compounds in this library to displacenorephinephrine ligands in vitro was determined by the methods of Galliet al (J. Exp. Biol. 198:2197, 1995) using desipramine (IC₅₀=920 nM) asa reference compound. The displacement of dopamine, and serotoninligands in vitro was determined by the methods of Gu et al (J. Biol.Chem. 269; 7124, 1994) using GBR-12909 (IC₅₀(DA uptake)=490 nM, IC₅₀(5-HT uptake)=110 nM) as a reference compound. The test compounds weretested at 1 μM (n=1) for dopamine (DA) uptake, norephinephrine (NE)uptake, and serotonin (5-HT) uptake. The table is a representation oftheir % inhibition at that test concentration.

Uptake Profile (% inhibition @ 1 μM) DA 5-HT Well # uptake NE uptakeuptake A1 90 87 49 A2 91 61 56 A3 86 78 59 A4 91 67 42 A5 96 64 73 A6 9094 44 A7 92 70 60 A8 90 60 32 A9 93 89 48 A10 93 71 56 A11 74 81 53 A1284 51 31 B1 88 88 48 B2 93 68 60 B3 85 85 52 B4 89 73 64 B5 97 68 77 B688 73 59 B7 94 60 59 B8 92 69 60 B9 74 71 60 B10 92 69 78 B11 78 68 75B12 76 60 62 C1 79 71 42 C2 90 52 54 C3 90 72 58 C4 85 61 57 C5 97 67 77C6 86 64 55 C7 91 66 53 C8 87 55 57 C9 91 75 59 C10 95 74 86 C11 61 5778 C12 78 66 68 D1 67 54 30 D2 78 57 55 D3 71 76 51 D4 86 61 46 D5 87 5773 D6 70 65 29 D7 83 61 54 D8 83 58 68 D9 76 69 59 D10 81 61 92 D11 5254 39 D12 68 52 76 E1 75 67 55 E2 86 47 63 E3 83 70 65 E4 86 55 67 E5 9765 75 E6 81 77 67 E7 86 72 56 E8 92 53 73 E9 85 57 71 E10 92 54 73 E1163 64 60 E12 80 50 83 F1 76 70 64 F2 77 56 51 F3 80 84 64 F4 87 66 59 F590 65 67 F6 77 68 70 F7 74 60 70 F8 87 60 82 F9 85 65 56 F10 84 46 52F11 89 56 57 F12 77 49 75 G1 88 82 78 G2 97 64 51 G3 90 77 75 G4 99 7478 G5 93 62 62 G6 91 85 74 G7 94 68 70 G8 99 87 74 G9 96 91 73 G10 95 8565 G11 89 79 81 G12 89 64 73 H1 65 65 39 H2 86 59 61 H3 77 77 54 H4 8970 64 H5 87 69 80 H6 81 72 53 H7 82 44 52 H8 86 55 67 H9 87 65 56 H10 8956 74 H11 85 60 47 H12 72 43 45

Example 100 Synthesis of1-(4-Chloro-phenyl)-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone

A solution of 3-(4-trifluoromethyl-phenoxymethyl)-piperidine (0.189mmol, 49 mg), 2-bromo-4′-chloroacetophenone (1.5 equiv, 0.284 mmol, 66mg) and K₂CO₃ (2.0 equiv, 0.378 mmol, 52 mg) in CH₃CN (1 mL) was heatedto 60° C. and stirred for 12 h. The reaction mixture was quenched withH₂O (10 mL) and then extracted with EtOAc (2×15 mL). The combinedorganics were dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm,5:1 hexane-EtOAc) provided 135 (37 mg, 78 mg theoretical, 47%) as acolorless oil: R_(f) 0.38 (SiO₂, 5:1 hexane-EtOAc); LRMS m/z 412 (M⁺+1,C₂₀H₂₁ClF₃NO₂, requires 412).

Example 101 Synthesis of1-(4-Chloro-phenyl)-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A solution of 135 (0.0898 mmol, 37 mg) in CH₃OH (400 μL) was treatedwith NaBH₄ (3.0 equiv, 0.269 mmol, 10 mg) at 0° C. The reaction mixturewas allowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (5 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(PTLC, SiO₂, 20 cm×20 cm, 1 mm, 6:1 Hexane-EtOAc) provided 136 (23 mg,37 mg theoretical, 62%) as a colorless oil: R_(f) 0.32 (SiO₂, 6:1Hexane-EtOAc); LRMS m/z 414 (M⁺+1, C₂₁H₂₃ClF₃NO₂, requires 414).

Example 102 Synthesis of 3-Styryl-piperidine-1-carboxylic Acid BenzylEster

A solution of Ph₃PCH₂Ph⁺C⁻ (1.5 equiv, 1.82 mmol, 708 mg) in THF (4 mL)was treated with nBuLi (1.5 equiv, 1.6M, 1.82 mmol, 1.14 mL) at −78° C.The solution was warmed to 0° C. for 30 min and then cooled again to−78° C. A solution of CBZ-piperidine-3-carbaldehyde (1.21 mmol, 300 mg)in THF (2 mL) was added to the above reaction mixture at −78° C. Thereaction stirred for 12 h. The reaction mixture was quenched with 10%HCl (10 mL) and then extracted with EtOAc (2×25 mL). The combinedorganics were dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (Isco Combi-Flash, 110 gcartridge, 9:1 Hexane-EtOAc) provided 137 (328 mg, 388 mg theoretical,85%) as a colorless oil: R_(f) 0.41 (SiO₂, 6:1 hexane-EtOAc); LRMS m/z322 (M⁺+1, C₂₁H₂₃NO₂, requires 322).

Example 103 Synthesis of 3-Phenethyl-piperidine

A solution of 137 (0.815 mmol, 262 mg) in CH₃OH (10 mL) was treated with30% PdC (50 mg) and H₂ (Parr Hydrogenator, 65 psi). The reaction wasshaken for 4 h. The reaction mixture was filtered through celite, andthe solvents were removed in vacuo to provide 138 (154 mg, 154 mgtheoretical, quantitative) as a colorless oil: LRMS m/z 190 (M⁺+1,C₁₃H₁₉N, requires 190).

Example 104 Synthesis of1-(4-Chloro-phenyl)-2-(3-phenethyl-piperidin-1-yl-ethanone

A solution of 138 (0.481 mmol, 91 mg), 2-bromo-4′-chloroacetophenone(1.5 equiv, 0.722 mmol, 169 mg) and K₂CO₃ (2.0 equiv, 0.962 mmol, 133mg) in CH₃CN (1 mL) was heated to 60° C. and stirred for 12 h. Thereaction mixture was quenched with H₂O (10 μL) and then extracted withEtOAc (2×25 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(PTLC, SiO₂, 20 cm×20 cm, 1 mm, 5:1 hexane-EtOAc) provided 139 (139 mg,164 mg theoretical, 85%) as a colorless oil: R_(f) 0.37 (SiO₂, 5:1hexane-EtOAc); LRMS m/z 342 (M⁺+1, C₂₁H₂₄ClNO, requires 342).

Example 105 Synthesis of1-(4-Chloro-phenyl)-2-(3-phenethyl-piperidin-1-yl)-ethanol

A solution of 139 (0.407 mmol, 139 mg) in CH₃OH (1 mL) was treated withNaBH₄ (3.0 equiv, 1.22 mmol, 46 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (5 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(PTLC, SiO₂, 20 cm×20 cm, 1 mm, 3:1 Hexane-EtOAc) provided 140 (74 mg,140 mg theoretical, 53%) as a colorless oil: R_(f) 0.36 (SiO₂, 3:1Hexane-EtOAc); LRMS m/z 344 (M⁺+1, C₂₁H₂₆ClNO, requires 344).

Example 106 Separation of 140 into its Four Constituent Diastereomers,141, 142, 143 and 144

140 was dissolved in 90:10 hexane (0.2% DEA) and isopropanol at aconcentration of 90 mg/mL. The compounds were separated on a ChiralpakAD column using the same solvent system as above providing the followingretention times: 141 (20.5 min), 142 (24.1 min), 143 (29.4 min) and 144(60.1 min). Syntheses starting with chiral R and S Ethyl nipecotatetartrates confirmed the stereochemistry at the 3-position of thepiperidine ring. The resulting diastereomers were separated utilizingthe same conditions.

Example 107 Synthesis of (S)-3-Styryl-piperidine-1-carboxylic AcidBenzyl Ester

A solution of Ph₃PCH₂Ph⁺Cl⁻ (1.5 equiv, 5.12 mmol, 1.99 g) in THF (10mL) was treated with nBuLi (1.5 equiv, 1.6M, 3.25 mmol, 2.04 mL) at −78°C. The solution was warmed to 0° C. for 30 min and then cooled again to−78° C. A solution of CBZ-piperidine-3-carboxaldehyde (3.41 mmol, 844mg) in THF (5 mL) was added to the above reaction mixture at −78° C. Thereaction stirred for 12 h. The reaction mixture was quenched with 10%HCl (20 mL) and then extracted with EtOAc (2×50 mL). The combinedorganics were dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (Isco Combi-Flash, 35 gcartridge, 9:1 Hexane-EtOAc) provided the olefin (647 mg, 1.10 mgtheoretical, 59%) as a colorless oil: R_(f) 0.41 (SiO₂, 6:1hexane-EtOAc); LRMS m/z 322 (M⁺+1, C₂₁H₂₃NO₂, requires 322).

Example 108 Synthesis of (R)-3-Phenethyl-piperidine

A solution of the olefin (0.0638 mol, 20.51 g) in CH₃OH (120 mL) wastreated 30% Pd—C (200 mg) and H₂ (Parr Hydrogenator, 65 psi). Thereaction was shaken for 4 h. The reaction mixture was filtered throughcelite, and the solvents were removed in vacuo to provide3-phenethyl-piperidine (12.08 g, 12.08 g theoretical, quantitative) as acolorless oil: LRMS m/z 190 (M⁺+1, C₁₃H₁₉N, requires 190).

Example 109 Synthesis of(R)-1-(4-Chloro-phenyl)-2-(3-phenethyl-piperidin-1-yl)-ethanone

A solution of 3-phenethyl piperidine (0.0158 mol, 3.00 g),2-bromo-4′-chloroacetophenone (1.0 equiv, 0.0158 mmol, 3.69 g) and KF(50% wt on celite) (8.0 equiv, 0.127 mol, 14.73 g) in CH₃CN (50 mL) wasstirred at rt for 12 h. The reaction mixture was then filtered, and thesolvents were removed in vacuo. Chromatography (Isco Combi-Flash, 35 gcartridge, 7:1 Hexane-EtOAc) provided 145 (2.88 g, 5.40 g theoretical,53%) as a colorless oil: R_(f) 0.37 (SiO₂, 5:1 hexane-EtOAc); LRMS m/z342 (M⁺+1, C₂₁H₂₄ClNO, requires 342).

Example 110 Synthesis of1-(4-Chloro-phenyl)-2-(3-phenethyl-piperidin-1-yl)-ethanol

A solution of 145 (8.16 mmol, 2.79 g) in CH₃OH (40 mL) was treated withNaBH₄ (2.0 equiv, 16.32 mmol, 617 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (100 mL) and extracted with EtOAc(2×100 mL). The combined organics were dried with NaCl_(4(sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 110 g cartridge, CH₂Cl₂ with 2% CH₃OH) provided 143and 144 as a mixture of diastereomers (1.91 g, 2.81 g theoretical, 68%)as a colorless oil: R_(f) 0.36 (SiO₂, 3:1 Hexane-EtOAc); LRMS m/z 344(M⁺+1, C₂₁H₂₆ClNO, requires 344). 143 and 144 were separated on aChiralpak AD column using 85:15 hexane (0.2% DEA) and isopropanol as theeluent. The retention times are as follow: 143 (30.35 min) and 144(65.32 min).

Example 111 Synthesis of (S)-3-Styryl-piperidine-1-carboxylic AcidBenzyl Ester

A solution of Ph₃PCH₂Ph⁺C⁻ (3.0 equiv, 28.05 mmol, 10.90 g) in THF (20mL) was treated with nBuLi (3.0 equiv, 2.5M, 28.05 mol, 11.2 mL) at −78°C. The solution was warmed to 0° C. for 30 min and then cooled again to−78° C. A solution of S-piperidine carboxaldehyde (9.35 mmol, 2.31 g) inTHF (20 mL) was added to the above reaction mixture at −78° C. Thereaction stirred for 12 h. The reaction mixture was quenched with 10%HCl (10 mL) and then extracted with EtOAc (2×25 mL). The combinedorganics were dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (Isco Combi-Flash, 110 gcartridge, 9:1 Hexane-EtOAc) provided the olefin (1.36 g, 3.01 gtheoretical, 45%) as a colorless oil: R_(f) 0.41 (SiO₂, 6:1hexane-EtOAc); LRMS m/z 322 (M⁺+1, C₂₁H₂₃NO₂, requires 322).

Example 112 Synthesis of (S)-3-Phenethyl-piperidine

A solution of the olefin (4.23 mmol, 1.36 mg) in CH₃OH (10 mL) wastreated 30% Pd—C (50 mg) and H₂ (Parr Hydrogenator, 65 psi). Thereaction was shaken for 4 h. The reaction mixture was filtered throughcelite, and the solvents were removed in vacuo to provide3-phenethyl-piperidine (801 mg, 801 mg theoretical, quantitative) as acolorless oil: LRMS m/z 190 (M⁺+1, C₁₃H₁₉N, requires 190).

Example 113 Synthesis of(S)-1-(4-Chloro-phenyl)-2-(3-phenethyl-piperidin-1-yl)-ethanone

A solution of (S)-3-phenethyl-piperidine (4.23 mmol, 801 mg),2-bromo-4′-chloroacetophenone (1.5 equiv, 6.35 mmol, 1.48 g) and K₂CO₃(2.0 equiv, 8.46 mmol, 1.17 g) in CH₃CN (10 mL) was heated to 60° C. andstirred for 12 h. The reaction mixture was quenched with H₂O (25 mL) andthen extracted with EtOAc (2×50 mL). The combined organics were driedwith NaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed in vacuoand chromatography (Isco Combi-Flash, 110 g cartridge, 9:1 Hexane-EtOAc)provided the desired product (0.556 mg, 1.45 mg theoretical, 38%) as acolorless oil: R_(f) 0.37 (SiO₂, 5:1 hexane-EtOAc); LRMS m/z 342 (M⁺+1,C₂₁H₂₄ClNO, requires 342).

Example 114 Synthesis of1-(4-Chloro-phenyl)-2-(3-phenethyl-piperidin-1-yl)-ethanol

A solution of the ketone (1.63 mmol, 556 mg) in CH₃OH (5 mL) was treatedwith NaBH₄ (3.0 equiv, 4.89 mmol, 185 mg) at 0° C. The reaction mixturewas allowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (20 mL) and extracted with EtOAc(2×30 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 35 g cartridge, 1:1 Hexane-EtOAc) provided 141 and142 as a mixture of diastereomers (366 mg, 561 mg theoretical, 65%) as acolorless oil: R_(f) 0.36 (SiO₂, 3:1 Hexane-EtOAc); LRMS m/z 344 (M⁺+1,C₂₁H₂₆ClNO, requires 344). 141 and 142 were separated on a Chiralpak ADcolumn using 85:15 hexane (0.2% DEA) and isopropanol as the eluent. Theretention times are as follow: 141 (20.19 min) and 142 (23.22 min).

Example 115 Synthesis of[1-(4-Chloro-phenyl)-cyclobutyl]-(3-phenethyl-piperidin-1-yl)-methanone

A solution of 3-phenethyl piperidine (0.449 mmol, 85 mg) and1-(4-chloro-phenyl)-cyclobutanecarboxylic acid (1.5 equiv, 0.674 mmol,142 mg) in CH₂Cl₂ (1 mL) was treated with PyBrOP (1.5 equiv, 0.674 mmol,314 mg) and iPr₂Net (3.0 equiv, 1.35 mmol, 235 μL) at 0° C. The reactionmixture stirred for 12 h while warming to rt. The reaction mixture wasquenched with 10% HCl (10 mL) and then extracted with EtOAc (2×15 mL).The combined organics were washed with NaHCO_(3(sat)) and dried withNaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed in vacuo andchromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm, 6:1 hexane-EtOAc)provided 146 (114 mg, 171 mg theoretical, 67%) as a colorless oil: R_(f)0.36 (SiO₂, 6:1 hexane-EtOAc); LRMS m/z 382 (M⁺+1, C₂₄H₂₈ClNO, requires382).

Example 116 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-phenethyl-piperidine

A solution of 146 (0.262 mmol, 100 mg) in toluene (1 mL) at 0° C. wastreated with 3.0 M Red-Al (65% wt in toluene) (3.0 equiv, 0.815 mmol)under Ar. The reaction mixture stirred for 12 h and returned to 25° C.The reaction mixture was then cooled to 0° C., quenched with 10% aqueousNaOH and extracted with 3×EtOAc (25 mL). The organics were dried withNaCl_((sat)) and Na₂SO_(4(s)). The reaction mixture was purified bychromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm, 6:1 hexane-acetone) whichprovided 147 (80 mg, 96 mg theoretical, 83%) as a colorless oil: R_(f)0.48 (SiO₂, 6:1 hexane-acetone); LRMS m/z 369 (M⁺+1, C₂₄H₃₀ClN, requires369).

Example 117 Synthesis of3-[2-(4-Trifluoromethyl-phenyl)-vinyl]-piperidine-1-carboxylic AcidBenzyl Ester

A solution of the wittig salt (1.5 equiv, 12.03 mmol, 6.03 g) in THF (40mL) was treated with nBuLi (1.5 equiv, 2.5M, 12.03 mmol, 4.8 mL) at −78°C. The solution was warmed to 0° C. for 30 min and then cooled again to−78° C. A solution of piperidine-3-carbaldehyde (8.02 mmol, 1.98 g) inTHF (10 mL) was added to the above reaction mixture at −78° C. Thereaction stirred for 12 h. The reaction mixture was quenched with 10%HCl (20 mL) and then extracted with EtOAc (2×50 mL). The combinedorganics were dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (Isco Combi-Flash, 110 gcartridge, 9:1 Hexane-EtOAc) provided 148 (2.08 g, 3.21 g theoretical,67%) as a colorless oil: R_(f) 0.44 (SiO₂, 6:1 hexane-EtOAc); LRMS m/z390 (M⁺+1, C₂₂H₂₂F₃NO₂, requires 390).

Example 118 Synthesis of3-[2-(4-Trifluoromethyl-phenyl)-ethyl]-piperidine

A solution of 148 (5.34 mmol, 2.08 g) in CH₃OH (30 mL) was treated with30% Pd—C (500 mg) and H₂ (Parr Hydrogenator, 65 psi). The reaction wasshaken for 4 h. The reaction mixture was filtered through celite, andthe solvents were removed in vacuo to provide 149 (2.08 g, 2.08 gtheoretical, quantitative) as a colorless oil: LRMS m/z 258 (M⁺+1,C₁₄H₁₈F₃N, requires 258).

Example 119 Synthesis of1-(4-Chloro-phenyl)-2-{3-[2-(4-trifluoromethyl-phenyl)-ethyl]-piperidin-1-yl}-ethanone

A solution of 149 (0.777 mmol, 200 mg), 2-bromo-4′-chloroacetophenone(1.0 equiv, 0.777 mmol, 182 mg) and KF (50% wt on celite) (7.0 equiv,5.44 mol, 632 mg) in CH₃CN (5 mL) was stirred for 12 h. The reactionmixture was filtered, and the solvents were removed in vacuo.Chromatography (Isco Combi-Flash, 35 g cartridge, 2:1 Hexane-EtOAc)provided 150 (178 mg, 318 mg theoretical, 56%) as a colorless oil: R_(f)0.24 (SiO₂, 2:1 hexane-EtOAc); LRMS m/z 410 (M⁺+1, C₂₂H₂₃ClF₃NO,requires 410).

Example 120 Synthesis of1-(4-Chloro-phenyl)-2-{3-[2-(4-trifluoromethyl-phenyl)-ethyl]-piperidin-1-yl}-ethanol

A solution of 150 (0.398 mmol, 163 mg) in CH₃OH (2 mL) was treated withNaBH₄ (1.5 equiv, 0.597 mmol, 23 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (5 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 35 g cartridge, 1:1 Hexane-EtOAc) provided 151, 152,153, and 154 as a mixture of diastereomers (124 mg, 164 mg theoretical,76%) as a colorless oil: R_(f) 0.38 (SiO₂, 2:1 Hexane-EtOAc); LRMS m/z412 (M⁺1, C₂₂H₂₅ClF₃NO, requires 412).

Example 121 Separation of 151, 152, 153, and 154

The four diastereomers were dissolved in 90:10 hexane (0.2% DEA) andisopropanol at a concentration of 100 mg/mL. The compounds wereseparated on a Chiralpak AD column using 85:15 hexane (0.2% DEA) andisopropanol providing the following retention times: 151 (23.75 min),152 (23.75 min), 153 (29.27 min) and 154 (45.1 min). Since 151 and 152eluted as one peak, the compounds were separated using a Chiralpak ADcolumn using 90:10 methanol, acetonitrile (0.1% DEA) providing thefollowing retention times 151 (9.26 min), and 152 (10.68 min).

Example 122 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-{3-[2-(4-trifluoromethyl-phenyl)-ethyl]-piperidin-1-yl}-ethanone

A solution of 149 (0.411 mmol, 106 mg),2-chloro-1-[1-(4-chloro-phenyl)-cyclobutyl]-ethanone (1.0 equiv, 0.411mmol, 100 mg) and KF (50% wt on Celite) (7.0 equiv, 2.88 mol, 335 mg) inCH₃CN (3 mL) was stirred for 12 h. The reaction mixture was filtered,and the solvents were removed in vacuo. Chromatography (IscoCombi-Flash, 10 g cartridge, 2:1 Hexane-EtOAc) provided 155 (136 mg, 168mg theoretical, 81%) as a colorless oil: R_(f) 0.38 (SiO₂, 2:1hexane-EtOAc); LRMS m/z 465 (M⁺+1, C₂₆H₂₉ClF₃NO, requires 465).

Example 123 Spontaneous Locomotor Activity in Rats Animals

Male Sprague-Dawley rats, (Iffa Crédo, Saint-Germain/L′Arbresle,France), weighing 200-250 g at the beginning of the study, were used.

During acclimatization period, rats were housed, 2 or 3 per cage, inMakrolon type III cages, in the animal room (temperature: 20±2° C.,humidity: minimum 45%, air changes: >12 per hour, light/dark cycle of 12h/12 h [on at 7:00 A.M.]). Animals were allowed a minimum of 5 daysperiod before experiment for acclimatization.

Rats received food (TrouwNutrition, Vigny, France) and water (tap waterin water bottle) ad libitum. Rats were placed on a sawdust bedding intheir cages (Goldchips, Trouw Nutrition, Vigny, France). On the daybefore experiments, food was withdrawn to have animals fasted overnight.

Preparation of the Test Substance(S) Suspension and of the ReferenceCompound

On the day of experiment, test item(s) were solubilized in 5% dextrose(w/v)/polyethylene glycol (PEG) 400 (4:1 v/v).

Administrations

All test compounds were administered at 20 mg/kg as a single i.p. dose.

Locomotor Activity Measurements

Twenty, 60 and 120 minutes after administration, rats were placed in aplastic box 30×30 cm in a room with low light intensity (maximum 50lux). Locomotor activity was determined during 20 min periods using avideo image analyzer (Videotrack, View Point, France). Number ofoccurrences, distance and duration of fast and slow movements, number ofoccurrences and duration of periods of inactivity and number of rearswere measured.

Results

Rats treated with compounds 114, 115, 113, and 110 exhibit a significantincrease in locomotor activity compared to control animals at 60 minutesafter i.p. administration at a dose of 20 mg/kg.

Example 124 Acute Toxicity Assessments

An in vivo evaluation was carried out to determine the maximum tolerateddose of numerous test compounds in two animal species (mouse and rat).The compounds were administered i.v. and the animals were then observedfor 72 h. Compounds 114, 115, 113, 110, 124, 125, 129, and 130 weresolubilized in 5% dextrose (w/v)/polyethylene glycol 400 (4:1 v/v).Compounds 126, 127, 131, and 132 were solubilized in 10%hydroxypropyl-β-cyclodextrin (w/v).

Compounds 114, 115, 113, 110, 125, 127, 131, 130, and 132 administeredat 30 mg/kg i.v. were well tolerated by the animals and did not causeany mortality after 72 h in mice and rats. Compound 129 administeredi.v. was well tolerated by mice and rats after 72 h at 20 mg/kg and 30mg/kg respectively. Compounds 124 and 126 administered at 10 mg/kg i.v.were well tolerated by the animals and did not cause any mortality after72 h in mice and rats.

Compounds 143 and 144 administered at 5 mg/kg i.v. were tolerated by theanimals and did not cause any mortality after 72 h in mice and rats.

Example 125 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-94-trifluoromethyl-phenoxy)-piperidine

To 3-hydroxypiperidine hydrochloride (1.0 g, 7.3 mmol) in DCM (40 mL)was added 1-(4-chlorophenyl)-1-cyclobutane carboxylic acid (2.3 g, 10.9mmol) and iPr₂NEt (6.3 mL, 36.3 mmol) followed by PyBroP (5.1 g, 10.9mmol). The resulting solution was allowed to stir overnight at roomtemperature before diluting with ethyl acetate and quenching with 10%KOH. The layers were separated and the aqueous layer further washed withethyl acetate. The combined organic layers were then dried (MgSO₄),filtered and concentrated in vacuo. The resulting residue was purifiedby flash column chromatography using 40% ethyl acetate/petroleum etherto provide the desired amide 156 (1.74 g, 82%).

To a solution of 156 (500 mg, 1.7 mmol) in DMF (10 mL) at roomtemperature was added NaH (205 mg, 60% wt., 5.1 mmol) and the suspensionallowed to heat to 70° C. After stirring at this temperature for 40minutes, 4-fluorobenzenetrifluoride was added and the reaction allowedto continue at this temperature for two and one half hours before addingan additional portion of 4-fluorobenzenetrifluoride (0.10 mL) andstirring for an additional hour. The reaction mixture was then dilutedwith ethyl acetate and quenched by the addition of brine. The organiclayer was separated and the aqueous layer further washed with ethylacetate. The combined organic extracts were then dried (MgSO₄), filteredand concentrated in vacuo. The resulting residue was purified by flashcolumn chromatography using 20% ethyl acetate/petroleum ether to providethe desired ether 157 (246 mg, 33%). LRMS calculated for C₂₃H₂₃ClF₃NO₂437.14, found (M+) 438.60.

To amide 157 (100 mg, 0.23 mmol) in toluene (1 mL) was cautiously addedRed-Al (0.24 mL, 0.80 mmol). The resulting solution was allowed to stirat room temperature for one hour before adding an additional portion ofRed-Al (0.10 mL, 0.34 mmol) and stirring at room temperature overnight.The reaction was then diluted with ethyl acetate and quenched with 10%aqueous KOH. The layers were separated and the aqueous layer furtherwashed with ethyl acetate. The combined organic layers were then dried(MgSO₄), filtered and concentrated in vacuo. The resulting residue waspurified by flash column chromatography using 1% 2M NH₃ in EtOH/DCM toprovide the desired amine 158 (69 mg, 71%). LRMS calculated forC₂₃H₂₅ClF₃NO 423.16, found (M+) 424.31. ¹H NMR (300 MHz, CDCl₃): 7.50(d, J=8.9 Hz, 2H), 7.24 (d, J=8.4 Hz, 2H), 7.08 (d, J=8.3 Hz, 2H), 6.71(d, J=8.5 Hz, 2H), 4.06-4.16 (m, 1H), 2.71-2.76 (m, 1H), 2.53-2.58 (m,2H), 2.42-2.46 (m, M1), 1.92-2.25 (m, 8H), 1.75-1.88 (m, 1H), 1.60-1.68(m, 1H), 1.41-1.56 (m, 1H), 1.25-1.38 (m, 1H). ¹³C NMR (75 MHz, CDCl₃):160.0, 148.2, 130.9, 127.8, 127.6, 126.9, 122.6 (m), 119.1, 115.2, 73.1,67.8, 59.2, 55.6, 47.0, 31.6, 31.2, 29.8, 23.7, 15.9.

Example 126 Synthesis of1-[4-(4-Chloro-phenyl)-tetrahydro-pyran-4-ylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)piperidine

Into a round-bottom flask under argon fitted with an addition funnel andthermometer was added anhydrous dimethylsulfoxide (60 mL) and sodiumhydride (1.17 g, 48.8 mmol, 95%). Then a solution of(4-chlorophenyl)acetonitrile (3.37 g, 22.2 mmol) and 2-bromoethyl ether(90%, 3.41 mL, 24.4 mmol) in diethyl ether (15 mL) was added slowly,while maintaining the reaction temperature at 20-30° C. The reactionmixture was maintained at room temperature for overnight. The reactionmixture was carefully quenched with water (50 mL) and then extractedwith hexane (3×100 mL). The organic extracts were combined, washed withwater (3×75 mL), brine (50 mL), dried over anhydrous magnesium sulfate,filtered, and concentrated to give a pale yellow oil. The oil waspurified by column chromatography on silica gel usinghexane/ethyl:acetate (80:20) to give 4.7 g of4-(4-chlorophenyl)tetrahydropyran-4-carbonitrile, as a colorless oil. ¹HNMR (CDCl₃, 300 MHz): δ 2.00-2.15 (m, 4H); 3.89 (dt, 2H, J₁=12.6 Hz,J₂=3 Hz); 4.05-4.11 (m, 2H); 7.37-7.45 (m, 4H); ¹³C NMR (CDCl₃, 300MHz): δ 36.66 (2C), 41.51, 64.98 (2C), 121.40, 127.03, 129.37, 134.31,138.46.

Into a round-bottom flask was added4-(4-chlorophenyl)tetrahydropyran-4-carbonitrile (0.442 g, 2 mmol),bis-(hydroxyethyl)ether (6 mL), and potassium hydroxide (0.337 g, 5.96mmol). The reaction mixture was heated at 215° C. for 3 h. The reactionmixture was allowed to cool to room temperature, carefully quenched withwater (20 mL) and then washed with diethyl ether (2×20 mL). The aqeuouslayer was made acidic with the addition of concentrated HCl. The aqueouslayer was extracted with diethyl ether (2×20 mL). These extracts werecombined, washed with brine (15 mL), dried over anhydrous sodiumsulfate, filtered, and concentrated to give 0.32 g of4-(4-chlorophenyl)tetrahydropyran-4-carboxylic acid, as a tan solid. ¹HNMR (d₄-methanol, 300 MHz): δ 1.86-1.96 (m, 2H); 2.45-2.51 (m, 2H);3.56-3.65 (m, 2H); 3.86-3.93 (m, 2H); 7.33-7.44 (m, 4H); ¹³C NMR(d₄-methanol, 300 MHz): δ 35.62 (2C), 66.76 (2C), 128.79, 129.77,134.22, 143.34, 177.29.

Into a round-bottom flask was added3-(4-trifluoromethylphenoxymethyl)piperidine (29.8 mg, 0.115 mmol),dichloromethane (0.5 mL), 4-(4-chlorophenyl)tetrahydropyran-4-carboxylicacid (30.4 mg, 0.127 mmol), diisopropylethylamine (0.0442 mL, 0.254mmol), and bromotris(dimethylamino)phosphonium hexafluorophosphate(0.049 g, 0.127 mmol). The reaction mixture was stirred at roomtemperature overnight. The reaction mixture was diluted with 5% aqueoushydrochloric acid (10 mL) and then extracted with ethyl acetate (2×20mL). The extracts were combined, washed with brine (7 mL), dried overanhydrous sodium sulfate, filtered, and concentrated to give a colorlessoil. The oil was purified by column chromatography on silica gel usinghexane/ethyl acetate (2:1) to give 30.8 mg of the amide, as a colorlessoil.

Into a round-bottom flask was added the amide (30.8 mg, 0.064 mmol),tetrahydrofuran (2 mL), lithium aluminum hydride (0.160 mL of a 1M THFsolution, 0.160 mmol). The reaction mixture was heated at reflux for 1 hand then allowed to cool to room temperature. The reaction mixture wascarefully quenched with 2.5% aqueous sodium potassium tartrate (10 mL)and then extracted with ethyl acetate (2×10 mL). The extracts werecombined, washed with brine (5 mL), dried over anhydrous sodium sulfate,filtered, and concentrated to give a colorless oil. The oil was purifiedby column chromatography on silica gel using hexane/ethyl acetate/2Nammonia in ethanol (85:14:1) to give 17.8 mg of 159, as a colorless oil.¹H NMR (CDCl₃, 300 MHz): δ 1.02-1.10 (m, 1H); 1.39-1.66 (m, 3H);1.82-1.94 (m, 4H); 2.04-2.12 (m, 3H); 2.25-2.41 (m, 4H); 3.47-3.53 (m,2H); 3.68-3.77 (m, 4H); 6.89 (d, 2H, J=8.4 Hz); 7.23-7.31 (m, 4H); 7.54(d, 2H, J=8.4 Hz); ¹³C NMR (CDCl₃, 300 MHz): δ 25.09, 26.74, 34.10,36.45, 41.34, 56.87, 59.41, 64.41, 70.27, 70.85, 114.56, 122.63, 122.90,127.03, 128.43, 128.93, 131.83, 143.53, 161.67.

Example 127 Synthesis of (R) and (S)2-bromo-1-[1-(4-chloro-phenyl)-cyclobutyl]-ethanol

The methyl ketone was prepared from the corresponding nitrile asfollows. To a solution of the carbonitrile (40 g) in 100 mL toluenesolution was added 200 mL MeMgBr ether solution (3.0 M, 3 eq.). Thereaction mixture was heated to boiling under nitrogen with a 95° C. oilbath to distill away the ether solvent and subsequently kept under thattemperature overnight. The reaction mixture was cooled to roomtemperature and then poured into a second flask containing 500 mL water.It was acidified by the addition of 5 M hydrochloric acid (500 mL) andthe mixture was brought to reflux for 2 h. The product was extractedinto ether (3×200 mL) and the organic layers were combined and driedwith Na₂SO₄. The solvent was then evaporated to supply pure methylketone (40 g, 92%). ¹H NMR (CDCl₃, 300 MHz): δ (ppm) 7.36 (d, 2H), 7.19(d, 2H), 2.72-2.81 (m, 2H), 2.35-2.45 (m, 2H), 1.95 (s, 3H), 1.86-1.94(m, 2H); ¹³C NMR (CDCl₃, 300 MHz): δ (ppm) 208.3, 141.9, 132.9, 129.1,127.9, 59.0, 30.7, 24.6, 16.1.

The bromo ketone was prepared as follows. A mixture of methyl ketone(21.2 g, 0.102 mol) and methanol (80 mL) was cooled to 5° C. and aceticacid containing 30% HBr (0.8 mL) was added. Bromine (5 mL, 0.097 mol)was then added dropwise to this solution over a 20 min period while thetemperature was maintained at 5° C. The reaction was kept at thattemperature overnight. The reaction mixture was then poured into 250 mLwater and the product was extracted into ether (3×250 mL). The organiclayers were combined and dried with Na₂SO₄. The solvent was evaporatedto supply pure bromo ketone (28 g, 96%), ¹H NMR (CDCl₃, 300 MHz): δ(ppm) 7.40 (d, 2H), 7.22 (d, 2H), 3.90 (s, 2H), 2.51-2.90 (m, 2H),2.04-2.45 (m, 2H), 1.95-2.04 (m, 2H); ¹³C NMR (CDCl₃, 300 MHz): δ (ppm)201.2, 140.6, 133.6, 129.4, 128.5, 57.8, 31.7, 31.4, 16.3.

Into a 500 mL three-necked flask, equipped with a magnetic stir bar, anitrogen inlet, was charged with 150 mL anhydrous THF,(R)-2-methyl-CBS-oxazaborolidine (1.0 M in toluene, 4.35 mL, 4.35 mmol)and borane-methyl sulfide (2.0 M in THF, 2.2 mL, 4.4 mmol). The reactionflask was cooled to 0° C. A solution of the bromo ketone (25 g, 87 mmol)in anhydrous THF (50 mL) and more borane-methyl sulfide (2.0 M in THF,28.2 mL, 56.4 mmol) were added simultaneously over a period of 2 h whilethe reaction was maintained at 0° C. Following the addition, thereaction was warmed up to room temperature and stirred for 10 h. Thereaction was cooled to 0° C. again and MeOH (25 mL) was carefully added(gas evolution!). The reaction mixture concentrated in vacuo (Me₂S wastrapped and oxidized with household bleach) and the residue dissolved intoluene (250 mL). The solution was washed with H₂SO₄ (0.2 M, 3×100 mL)and water (3×100 mL), dried (Na₂SO₄) and concentrated. The product waspurified by column chromatography on silica gel using hexane/ethylacetate (95:5) to give the (R)-bromo alcohol (23.1 g, 92%) as colorlessoil, [α]²⁵ _(D)=−7.98 (c=1.19, CHCl₃), 99.5% ee; ¹H NMR (CDCl₃, 300MHz): δ (ppm) 7.32 (d, 2H), 7.14 (d, 2H), 4.09-4.13 (dd, 1H), 3.43-3.47(dd, 1H), 2.86-2.93 (t, 1H), 2.59-2.65 (m, 1H), 2.28-2.44 (m, 3H),2.01-2.10 (m, 1H), 1.90-1.92 (m, 1H); ¹³C NMR (CDCl₃, 300 MHz): δ (ppm)143.5, 132.4, 128.8, 128.4, 50.1, 37.4, 31.2, 30.6, 16.2.

Into a 500 mL three-necked flask, equipped with a magnetic stir bar, anitrogen inlet, was charged with 150 ml anhydrous THF,(s)-2-methyl-CBS-oxazaborolidine (1.0 M in toluene, 4.35 mL, 4.35 mmol)and borane-methyl sulfide (2.0 M in THF, 2.2 mL, 4.4 mmol). The reactionflask was cooled to 0° C. A solution of the bromo ketone (25 g, 87 mmol)in anhydrous THF (50 mL) and more borane-methyl sulfide (2.0 M in THF,28.2 mL, 56.4 mmol) were added simultaneously over a period of 2 h whilethe reaction was maintained at 0° C. Following the addition, thereaction was warmed up to room temperature and stirred for 10 h. Thereaction was cooled to 0° C. again and MeOH (25 mL) was carefully added(gas evolution!). The reaction mixture concentrated in vacuo (Me₂S wastrapped and oxidized with household bleach) and the residue dissolved intoluene (250 mL). The solution was washed H₂SO₄ (0.2 M, 3×100 mL) andwater (3×100 mL), dried (Na₂SO₄) and concentrated. The product waspurified by column chromatography on silica gel using hexane/ethylacetate (95:5) to give the (S)-bromo alcohol (24.8 g, 99%) as colorlessoil, [α]²⁵ _(D)=+8.24 (c=0.85, CHCl₃), 99.2% ee; ¹H NMR (CDCl₃, 300MHz): δ ppm) 7.32 (d, 2H), 7.14 (d, 2H), 4.09-4.13 (dd, 1H), 3.43-3.47(dd, 1H), 2.86-2.93 (t, 1H), 2.59-2.65 (m, 1H), 2.28-2.44 (m, 3H),2.01-2.10 (m, 1H), 1.90-1.92 (m, 1H); ¹³C NMR (CDCl₃, 300 MHz): δ (ppm)143.5, 132.4, 128.8, 128.4, 50.0, 37.4, 31.2, 30.6, 16.1.

Example 128 Synthesis of 2R-2-[1-(4-Chloro-phenyl)-cyclobutyl]-oxirane

A 500 mL round bottom flask was charged with2R-bromo-1-[1-(4-chloro-phenyl)-cyclobutyl]-ethanol (4.86 g; 16.8 mmol),THF (100 mL), MeOH (100 mL) and potassium carbonate (4.63 g; 33.6 mmol).The reaction mixture was stirred at 20° C. for 3 hours and then dilutedwith hexanes, filtered and concentrated in vacuo. The crude material waspurified by flash chromatography (hexanes/EtOAc 97:3) to give pureproduct (3.0 g; 86% yield).

The enantiomeric epoxide (2S-2-[1-(4-Chloro-phenyl)cyclobutyl]-oxirane)was also prepared according to the procedure described above, using2S-bromo-1-[1-(4-chloro-phenyl)-cyclobutyl]-ethanol (5.0 g, 17.3 mmol),THF (100 mL), MeOH (100 mL), and potassium carbonate (4.77 g, 34.5mmol). The crude material was purified by flash chromatography(hexanes/EtOAc 97:3) to give pure product (2.85 g; 79% yield).

Example 129 Synthesis of1R-1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[(3S)-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A 25 mL RB flask was charged with amine (3.79 g; 14.6 mmol) and2R-2-[1-(4-Chloro-phenyl)-cyclobutyl]-oxirane (3.05 g; 14.6 mmol) andheated to 95° C. with stirring for 12 hours. The reaction mixture wascooled to 20° C. and the crude material was purified by flashchromatography (hexanes/EtOAc 1:1 w/5% 2.0 M NH₃ in EtOH) to give purematerial (5.79 g; 85% yield). The diastereomeric purity was determinedto be 96.9% de based on chiral HPLC analysis.

Example 130 Synthesis of1R-1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[(3R)-3-(4-trifluoromethyl-phenoxymethyl)-piperidine-1-yl]-ethanol

A 25 mL RB flask was charged with amine (1.0 g; 3.86 mmol) and2R-2-[1-(4-chloro-phenyl)cyclobutyl]-oxirane (0.8 g; 3.86 mmol), CH₃CN(4 mL) and heated to 95° C. with stirring for 5 hours. The reactionmixture was cooled to 20° C. and concentrated in vacuo. The crudematerial was purified via crystallization from hot methanol to yieldpure product (998 mg, 56%). The diastereomeric purity was determined tobe 100% de based on chiral HPLC analysis. ¹H (300 MHz, CDCl₃) δ 7.55 (d,2H, J=9 Hz), 7.30 (d, 2H, J=6.5 Hz), 7.13 (d, 2H, J=9 Hz), 6.94 (d, 2H,9 Hz), 3.94-3.76 (m, 4H), 2.95-1.55 (m, 16H); ¹³C (100 MHz, CDCl₃) δ161.7, 145.2, 131.7, 128.9, 128.1, 127.1, 114.6, 71.3, 71.1, 59.7, 59.0,53.0, 48.9, 36.8, 31.0, 29.3, 27.3, 25.0, 16.3; IR (NaCl, cm⁻¹): 3423,2942, 1617, 1521, 1334, 1260, 1108, 1068, 836; MH⁺ (468).

Example 131 Synthesis of1S-1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[(3S)-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A 25 mL RB flask was charged with amine (239 mg; 0.92 mmol) and2S-2-[1-(4-chloro-phenyl)-cyclobutyl]-oxirane (192 mg; 0.92 mmol),CH₃CN, and heated to 95° C. with stirring for 3 hours. The reactionmixture was cooled to 20° C. and concentrated. The crude material waspurified by flash chromatography (hexanes/EtOAc 1:1 w/5% 2.0 M NH₃ inEtOH) to give pure material (324 g; 74% yield, 5de: 99.6%). ¹H (300 MHz,CDCl₃) δ 7.55 (d, 2H, J=9 Hz), 7.30 (d, 2H, J=6.5 Hz), 7.13 (d, 2H, J=9Hz), 6.94 (d, 2H, 9 Hz), 3.94-3.76 (m, 4H), 2.95-1.55 (m, 16H); ¹³C (100MHz, CDCl₃) δ 161.7, 145.2, 131.7, 128.9, 128.1, 127.1, 114.6, 71.3,71.1, 59.7, 59.0, 53.0, 48.9, 36.8, 31.0, 29.3, 27.3, 25.0, 16.3; IR(NaCl, cm⁻¹): 3423, 2942, 1617, 1521, 1334, 1260, 1108, 1068, 836;MH+(468).

Example 132 Synthesis of1S-1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[(3R)-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A 25 mL RB flask was charged with amine (1.5 g; 5.79 mmol) and2S-2-[1-(4-chloro-phenyl)-cyclobutyl]-oxirane (1.2 g; 5.79 mmol) andheated to 95° C. with stirring for 5 hours. The reaction mixture wascooled to 20° C. and the crude material was purified by flashchromatography (hexanes/EtOAc 1:1 w/5% 2.0 M NH₃ in EtOH) to give purematerial (2.31 g; 85% yield). The diastereomeric purity was determinedto be 99.42% de based on chiral HPLC analysis. ¹H (300 MHz, CDCl₃) δ7.58 (m, 2H), 7.31 (m, 2H), 7.17 (m, 2H), 6.98 (m, 2H), 3.94-3.82 (m,3H), 3.11 (m, 2H), 2.60-1.63 (m, 15H); ¹³C (100 MHz, CDCl₃) δ 161.7,145.2, 131.8, 128.9, 28.1, 127.1, 114.6, 71.2, 59.6, 56.0, 49.0, 36.6,31.0, 29.1, 27.4, 25.3, 16.3; IR(NaCl, cm⁻¹): 3428, 2932, 1623, 1320,1259, 1155, 1105, 1061, 836; MH⁺ (467).

Example 133 Synthesis of1R-1-{2-[1-(4-Chloro-phenyl)-cyclobutyl]-2-methoxy-ethyl}-(3R)-3-(4-trifluoromethyl-phenoxymethyl)-piperidine

The alcohol (107.6 mg, 0.230 mmol) was dissolved in THF (2.5 mL). Methyliodide (0.072 mL, 1.15 mmol) and potassium tert-butoxide (39 mg, 0.345mmol) were added. The reaction continued stirring at RT and wasmonitored by HPLC. After completion the reaction mixture was dilutedwith water and the aqueous layer was extracted with EtOAc (3×, 20 mL).Combined organic layers were dried over Na₂SO₄ and concentrated. Thecrude material was purified using silica gel chromatography (80:16:4hexanes; EtOAc: 2M ammonia in EtOH) to yield pure product. MH⁺=481.

Example 134 Synthesis of1S-1-{2-[1-(Chloro-phenyl)-cyclobutyl]-2-methoxy-ethyl}-(3R)-3-(4-trifluoromethyl-phenoxymethyl)-piperidine

1S-1-{2-[1-(4-Chloro-phenyl)-cyclobutyl]-2-methoxy-ethyl}-(3R)-3-(4-trifluoromethyl-phenoxymethyl)-piperidinewas prepared according to the procedure in Example 133: alcohol (100 mg,0.21 mmol), potassium tert-butoxide (36 mg, 0.32 mmol), MeI (151.6 mg,1.07 mmol), THF (2.2 mL). The crude material was purified using silicagel chromatography (80:16:4 hexanes:EtOAc: 2M ammonia in EtOH) to yieldpure product. MH⁺=481.

Example 135 Synthesis of1R-1-{2-[1-(4-Chloro-phenyl)-cyclobutyl]-2-methoxy-ethyl}-(3S)-3-(4-trifluoromethyl-phenoxymethyl)-piperidine

1R-1-{2-[1-(4-Chloro-phenyl)-cyclobutyl]-2-methoxy-ethyl}-(38)-3-(4-trifluoromethyl-phenoxymethyl)-piperidinewas prepared according to the procedure in Example 133: alcohol (289 mg,0.62 mmol), potassium tert-butoxide (104 mg, 0.93 mmol), MeI (430 mg,3.0 mmol), THF (5 mL). The crude material was purified using silica gelchromatography (1:1 hexanes: EtOAc in 2M ammonia in EtOH) to yield pureproduct (256 mg, 86%).

Example 136 Synthesis of1S-1-{2-[1-(4-Chloro-phenyl)-cyclobutyl]-2-methoxy-ethyl}-(3S)-3-(4-trifluoromethyl-phenoxymethyl)-piperidine

1S-1-{2-[1-(4-Chloro-phenyl)-cyclobutyl]-2-methoxy-ethyl}-(3S)-3-(4-trifluoromethyl-phenoxymethyl)-piperidinewas prepared according to the procedure in Example 133: alcohol (200 mg,0.43 mmol), potassium tert-butoxide (78 mg, 0.64 mmol), MeI (303 mg, 2.1mmol), THF (5 mL). The crude material was purified using silica gelchromatography (1:1 hexanes: EtOAc in 2M ammonia in EtOH) to yield pureproduct (196 mg, 95%).

Example 137 Synthesis of[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-3-ol(167)

5-Benzyl-1-oxa-5-aza-spiro[2.5]octane

Sodium hydride (583 mg, 14.6 mmol) in DMSO (6 mL) was heated to 55 C for1 h. The reaction mixture is cooled to 0 C and Me₃SI (3 g, 14.62 mmol)dissolved in THF (9.8 mL) was added dropwise. 1-Benzyl-piperidine-3-one(1.5 g, 6.64 mmol) dissolved in DMSO (10 mL) was added 15 minutes later.After completion of addition the reaction proceeded at RT. After 30 min.the reaction was quenched with water. The aqueous layer was extractedwith hexanes. Combined organic layers were dried over Na₂SO₄ andconcentrated. The crude material was purified using silica gelchromatography (9:1 DCM:hexanes in 2 M ammonia in EtOH) to yield5-benzyl-1-oxa-5-aza-spiro[2.5]octane (960 mg, 71%). MH+(204).

1-Benzyl-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-3-ol

5-benzyl-1-oxa-5-aza-spiro[2.5]octane (960 mg, 4.74 mmol) dissolved indioxane (3 mL) was added dropwise to a hot (105 C) stirring solution ofNaOH (189 mg, 4.74 mmole), p-trifluoro-cresol (2.30 g, 14.2 mmol), anddioxane (3 mL). After completion of addition the reaction mixture wasstirred at 110 C for 6 h and at RT for 12 h. The reaction mixture wasdiluted with 10% NaOH and extracted with diethyl ether. Combined organiclayers were concentrated to yield a brown oil. The crude material waspurified using silica gel chromatography (4:1 hexane:EtOAc in 2M ammoniain EtOH) to yield1-benzyl-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-3-ol (100 mg,6%). MH+(365).

3-(4-Trifluoromethyl-phenoxymethyl)-piperidin-3-ol

The amine (100 mg 0.274 mmol) was dissolved in EtOH (3 mL). 10% Pd/Ccatalyst (37 mg) was added to the solution. The reaction mixture wasstirred under H₂ atmosphere at 50 psi for 5 h. The catalyst was removedvia filtration and the filtrate was concentrated to afford the desiredcompound as a yellow oil (75 mg, 100%).

[1-(4-Chloro-phenyl)-cyclobutyl]-[3-hydroxy-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-methanone

Into a round-bottom flask was added3-(4-trifluoromethyl-phenoxymethyl)-piperidin-3-ol (100 mg, 0.36 mmol),dichloromethane (S mL), diisopropylethylamine (0.190 mL, 1.08 mmol),1-(4-chloro-phenyl)-cyclobutanecarboxylic acid (113.4 mg, 0.54 mmole),and PyBroP (0.252 g, 0.54 mmol). The reaction mixture was stirred atroom temperature overnight. The reaction mixture was diluted with waterand then extracted with ethyl acetate (3×10 mL). The extracts werecombined, dried over anhydrous sodium sulfate, filtered, andconcentrated to give a yellow oil. The oil was purified by columnchromatography on silica gel using hexane/ethyl acetate (4:1) to give[1-(4-Chloro-phenyl)-cyclobutyl]-[3-hydroxy-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-methanone(18.7 mg, 11%). MH+ (468).

[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethyl-phenoxymethyl)-piperidin-3-ol

The amide (18.7 mg, 0.40 mmol) was dissolved in THF (1 mL) and cooled inan ice bath. LAH (1M in THF, 0.048 mL, 0.048 mmol) was added to thecooled stirring reaction mixture. After completion of addition thereaction stirred at RT. After 12 h the reaction mixture was quenchedwith water. The aqueous layer was extracted with EtOAc (3×5 mL). Thecombined organic layers were dried over Na₂SO₄ and concentrated. Thecrude material was purified using silica gel prep plate (4:1 hexaneEtOAc in 2M ammonia in EtOH) to yield the desired compound (167).MH+(454).

Example 138 Synthesis of1-[1-(2-Methoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

The starting acid 1-(2-methoxy-phenyl)-cyclobutanecarboxylic acid hasbeen described (S. L. Mnhzhoyan et al. Pharm. Chem., Eng. Translation,1980, 14 (2), 114-118). A mixture of this acid (145 mg, 0.70 mmol) andthionyl chloride (2 mL) was heated at reflux for 3 hr. The reactionmixture was concentrated in vacuo, diluted with THF (2 mL),reconcentrated, and residual solvent was removed by vaccuum. Thematerial was dissolved in 2 mL THF, cooled to 0° C., and treated withexcess diazomethane in ether (generated from 0.5 g1-methyl-3-nitro-1-nitrosoguanidine in 3 mL ether and 0.34 g NaOH in 3mL of water). The solution was stirred overnight at 0 C, and then HCl (1mL of a 4M solution in dioxane) was added and the mixture was kept atthat temperature for 1 hr. The solution was concentrated in vacuo andpurified on silica gel (9:1 ethyl acetate/hexane) to give2-chloro-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanone as a colorless oil(92 mg, 55%). Data for this chloroketone: MS 239 (M+1); ¹H NMR (300 MHz,CDCl₃): δ 6.85-7.4 (m, 4H), 3.89 and 3.86 (singlets, total 5H), 2.0-2.9(m, 6H).

2-Chloro-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanone (55 mg) in 2 mLacetonitrile was treated with freshly flamed-dried KF on Celite (200 mg)and 3-(4-trifluoromethyl-phenoxymethyl)-piperidine (66 mg, 1.1 equiv).The mixture was stirred overnight, diluted with THF (5 mL), filtered,concentrated in vacuo and purified on silica gel to give the desired1-[1-(2-methoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone(32 mg, 30%). Data for this amino ketone: MS 462 (M+1); ¹H NMR (300 MHz,CDCl₃) partial: δ 6.8-7.6 (m, 8H), 3.85-3.95 (m, 2H, methylene adjacentto aryl ether), 3.09 (s, 2H, methylene adjacent to keto and aminemoieties).

1-[1-(2-Methoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone(15 mg) was dissolved in 2 mL of dry methanol and solid sodiumborohydride (10 equiv.) was added in portions. Water (5 mL) was addedand the mixture was extracted with ether and the ether extracts wereconcentrated in vacuo and purified on silica gel to give the desired1-[1-(2-methoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol(9 mg, 60%). Data for this mixture of diastereomeric amino alcohols: MS464 (M+1); ¹H NMR (300 MHz, CDCl₃) partial: δ 6.8-7.6 (m, 8H), 3.85-3.95(m, 2H, methylene adjacent to aryl ether), the 63.09 singlet for thestarting material was absent.

Example 139 Synthesis of1-(1-benzo[1,3]dioxol-5-yl-cyclobutyl)-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A mixture of benzo[1,3]dioxol-5-yl-acetonitrile (1.60 g, 9.94 mmol) andthe dibromide (1.11 mL, 1.1 equiv.) in 10 mL of DMSO was added to amixture of NaH (1 g 60% suspension, 2.5 equiv.) in 30 mL of DMSO at roomtemperature. After 24 hours the reaction was quenched by addition of 50mL of pH 7.0 buffer solution, the mixture was extracted with ether, andthe ether extracts were concentrated in vacuo and purified on silica gel(85:15 hexane:ethyl acetate) to give the desired1-benzo[1,3]dioxol-5-yl-cyclobutanecarbonitrile (1.41 g, 71%). Data forthis nitrile: MS 202 (M+1); ¹H NMR (300 MHz, CDCl₃): δ 6.8-6.9 (m, 3H),5.98 (s, 2H), 2.0-2.85 (m, 6H); ¹³C NMR (75 MHz, CDCl₃): δ 148.5, 147.5,134.0, 124.7, 119.2, 106.5, 101.6, 40.2, 35.0, 17.2.

1-Benzo[1,3]dioxol-5-yl-cyclobutanecarbonitrile (1.2 g) was slurried inethylene glycol (10 mL) in a pressure tube and a large excess of KOH (1g) was added. The tube was sealed and heated at 180° C. for 24 hours,cooled, poured into 100 mL of water, acidified with 1N HCl, extractedwith ether, and the ether extracts were concentrated in vacuo andchromatographed on silica gel with an ethyl acetate:hexane:acetic acidmixture to give the desired1-benzo[1,3]dioxol-5-yl-cyclobutanecarboxylic acid (850 mg, 65%). Datafor this acid: MS 220 (M); ¹H NMR (300 MHz, CDCl₃): δ 11.4-11.8 (br. s,1H), 2.78-6.85 (m, 3H), 5.95 (s, 2H) 2.75-2.85 (m, 2H), 2.40-2.55 (m,2H), 1.8-2.2 (m, 2H); ¹³C NMR (75 MHz, CDCl₃): δ 182.9, 147.9, 146.7,137.2, 119.8, 108.2, 107.5, 101.3, 52.2, 32.6, 16.7.

1-Benzo[1,3]dioxol-5-yl-cyclobutanecarboxylic acid was converted to thecorresponding chloroketone following the same procedure used for theconversion of 1-(2-Methoxy-phenyl)-cyclobutanecarboxylic acid to give2-Chloro-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanone. 300 mg of1-benzo[1,3]dioxol-5-yl-cyclobutanecarboxylic acid thus provided 285 mg(83%) of 1-(1-Benzo[1,3]dioxol-5-yl-cyclobutyl)-2-chloro-ethanone. Datafor this chloride: ¹H NMR (300 MHz, CDCl₃): δ 6.7-6.85 (m, 3H), 5.98 (s,2H), 4.03 (s, 2H), 2.75-2.85 (m, 2H), 2.35-2.45 (m, 2H), 1.85-2.0 (m,2H); ¹³C NMR (75 MHz, CDCl₃): δ 201.9, 148.6, 147.1, 135.8, 119.8,108.9, 107.0, 101.6, 57.7, 45.4, 31.2, 16.2.

1-(1-Benzo[1,3]dioxol-5-yl-cyclobutyl)-2-chloro-ethanone was used forthe alkylation of 3-(4-Trifluoromethyl-phenoxymethyl)-piperidine by thesame method used for the alkylation of2-Chloro-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanone with this amine.100 mg of 1-(1-Benzo[1,3]dioxol-5-yl-cyclobutyl)-2-chloro-ethanone wasused to prepare 85 mg of1-(1-benzo[1,3]dioxol-5-yl-cyclobutyl)-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone(45%) by the KF-Celite method described in Example 138. Data for thisamino ketone: MS 476 (M+1); ¹H NMR (300 MHz, CDCl₃) partial: δ 6.7-7.7(m, 7H), 6.0 (s, 2H), 3.85-3.95 (m, 2H, methylene adjacent to arylether), 3.1 (s, 2H, methylene adjacent to keto and amine moieties).

1-(1-Benzo[1,3]dioxol-5-yl-cyclobutyl)-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone(15 mg) was dissolved in 2 mL of dry methanol and solid sodiumborohydride (10 equiv.) was added in portions. Water (5 mL) was addedand the mixture was extracted with ether and the ether extracts wereconcentrated in vacuo and purified on silica gel to give the desired1-(1-benzo[1,3]dioxol-5-yl-cyclobutyl)-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol(10 mg, 66%). Data for this mixture of diastereomeric amino alcohols: MS478 (M+1); ¹H NMR (300 MHz, CDCl₃) partial: δ 6.7-7.8 (m, 7H), 3.85-3.95(m, 2H, methylene adjacent to aryl ether), the δ 3.1 singlet for thestarting material was absent.

Example 140 Synthesis of2-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-1-[1-(4-trifluoromethyl-phenyl)-cyclobutyl]-ethanol

The starting nitrile,1-(4-Trifluoromethyl-phenyl)-cyclobutanecarbonitrile, has been described(Parke Davis & Co., U.S. Pat. No. 3,536,656; 1970 and Chemical Abstracts1970, 73, 109539). To the carbonitrile (2 g) in 5 mL of toluene in apressure tube was added 10 mL of 3M MeMgBr ether solution (3 equiv.).The tube was sealed and the reaction mixture was heated at 95° C. for 24hr, cooled to room temperature, poured into 50 mL of water, acidifiedwith 25 mL of 5 M HCl, and this solution was heated at 70° C. for 2 h,cooled, and extracted with ether. The extracts were dried, filtered,concentrated, and purified on silica gel to give pure 1-[1-(4trifluoromethyl-phenyl)-cyclobutyl]-ethanone (1.9 g, 88%). Data for thisketone: MS 242 (M); ¹H NMR (300 MHz, CDCl₃) δ 7.6 (d, 8.0 Hz, 2H), 7.4(d, 8.0 Hz, 2H), 2.75-2.85 (m, 2H), 2.35-2.5 (m, 2H), 1.8-2.0 (3H s and2H m overlapping); ¹³C NMR (75 MHz, CDCl₃): δ 207.7, 147.5, 129.5,126.9, 126.1, 125.9 (CF₃), 59.5, 30.9, 24.6, 16.1.

1-[1-(4-Trifluoromethyl-phenyl)-cyclobutyl]-ethanone (1.06 g) inmethanol (8 mL) was cooled to 0° C. Acetic acid containing 30% HBr (0.35mL) was added, and then precisely 1 molar equivalent of bromine wasadded dropwise. The reaction mixture was maintained at 0° C. overnight,poured into 20 mL of water, and extracted with ether. The extracts weredried, filtered, concentrated, and purified on silica gel to give pure2-bromo-1-[1-(4-trifluoromethyl-phenyl)-cyclobutyl]-ethanone (1.12 g,80%). Data for this bromoketone: MS 321 (M); ¹H NMR (300 MHz, CDCl₃) δ7.82 (d, 8 Hz, 2H), 7.39 (d, 8 Hz, 2H), 3.82 (s, 2H), 2.80-2.95 (m, 2H),2.45-2.60 (m, 2H), 1.90-2.10 (m, 2H).

2-Bromo-1-[1-(4-trifluoromethyl-phenyl)-cyclobutyl]-ethanone was usedfor the alkylation of 3-(4-Trifluoromethyl-phenoxymethyl)-piperidine bythe same method used for the alkylation of2-Chloro-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanone with this amine.100 mg of 2-Bromo-1-[1-(4-trifluoromethyl-phenyl)-cyclobutyl]-ethanonewas converted to 96 mg of2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-1-[1-(4-trifluoromethyl-phenyl)-cyclobutyl]-ethanone(62%) by the KF-Celite method described in Example 138. Data for thisamino ketone: MS 500 (M+1); ¹H NMR (300 MHz, CDCl₃): δ 7.61 (d, 8.3 Hz,2H), 7.52 (d, 8.7 Hz, 2H), 7.38 (d, 8.3 Hz, 2H), 6.91 (d, 8.7 Hz, 2H),3.75-3.85 (m, 2H, methylene adjacent to aryl ether), 3.06 (s, 2H),1.0-2.9 (various overlapping multiplets totaling 15H); ¹³C NMR (75 MHz,CDCl₃): 6207.0, 161.7, 147.3, 128.6, 127.8, 127.11, 127.06, 126.99,125.93, 125.88, 114.6, 71.2, 63.2, 58.3, 57.2, 54.2, 36.2, 31.2, 31.1,27.0, 24.8, 16.5.

2-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-1-[1-(4-trifluoromethyl-phenyl)-cyclobutyl]-ethanone(30 mg) was dissolved in 3 mL of dry methanol and solid sodiumborohydride (10 equiv.) was added in portions. Water (10 mL) was addedand the mixture was extracted with ether and the ether extracts wereconcentrated in vacuo and purified on silica gel to give the desired2-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-1-[1-(4-trifluoromethyl-phenyl)-cyclobutyl]-ethanol(21 mg, 67%). Data for this mixture of diastereomeric amino alcohols: MS502 (M+1); ¹H NMR (300 MHz, CDCl₃): δ 7.5-7.6 (m, 4H), 7.29-7.35 (m,2H), 6.89-6.96 (overlapping doublets, total 2H), 3.8-4.0 (m, 4H, 1 ofwhich is exchanged upon exposure to D₂O), 1.1-3.1 g (various overlappingmultiplets totaling 17H); ¹³C NMR (75 MHz, CDCl₃): δ 207 peak instarting ketone is absent. Most peaks are doubled (adjacent equalintensity peaks) due to diastereoisomerism: δ 161.7, 151.0, 128.6 &128.5, 128.0 & 127.8, 127.15 & 127.10, 126.45, 124.91 & 124.87, 123.30,122.86, 114.63, 71.2 & 71.1, 59.7 & 59.5, 58.99 & 58.97, 55.96, 49.31 &49.28, 36.8 & 36.6, 31.0, 29.5 & 29.4, 27.33 & 27.26, 25.24 & 25.03,16.42.

Example 141 Spontaneous Locomotor Activity in Rats

The effect of 124 and 126 on spontaneous locomotor activity in rats wasdetermined according to the procedures outlined by Silverman et al.(Motor Activity. In “Animal behavior in the laboratory”, Chapman andHall eds, London, p. 79-92, 1978) and Boissier et al. (Arch. Int.Pharmacodyn. 1965, 158, 212.)

Test items and test item vehicles were administered to maleSprague-Dawley rats (n=10) as a single i.p. dose (vehicle A=10%hydroxypropyl-β-cyclodextrin in water; vehicle B=4:1 5% aqueousdextrose:PEG 400). Twenty, 60 and 120 minutes following administration,rats were placed in a plastic box 30×30 cm in a room with low lightintensity (maximum 50 lux). Locomotor activity was determined during 20minute periods using video image analyzers. Images recorded with videocameras were digitalized and displacements of the center of gravity ofthe digital image spot were tracked and analyzed. When the speed ofdisplacement of the center of gravity of the spot was below 4.26 cm/sec,the movement was considered as inactivity. When this speed was between4.26 and 6.75 cm/sec, the movement was considered as a small movement.When this speed was above 6.75 cm/sec, the movement was considered as alarge movement. The number of occurrences, distance and duration of fastand slow movements, number of occurrences and duration of periods ofinactivity and number of rears were measured.

Results

Compounds 126 and 124, when dosed at 5 and 10 mg/kg, exhibit asignificant increase in locomotor activity compared to control animals.

Methyl- 126 126 124 124 phenidate, (5 mg/kg (10 mg/kg (5 mg/kg (10 mg/kg(10 mg/kg in Vehicle in Vehicle in Vehicle in Vehicle in Vehicle VehicleA Vehicle B A) A) B) B) B) Large  20 min:  20 min:  20 min:  20 min:  20min:  20 min:  20 min: Movement 303.7 ± 28.0 317.4 ± 15.5 287.0 ± 34.4364.7 ± 52.7  228.7 ± 22.5 391.5 ± 63.3  974.3 ± 125.5 Occurrences  60min:  60 min:  60 min:  60 min:  60 min:  60 min:  60 min: 104.9 ± 11.7 99.4 ± 19.7 204.4 ± 36.3 644.5 ± 127.3  74.3 ± 14.8 427.6 ± 95.8 423.9± 75.9 120 min: 120 min: 120 min: 120 min: 120 min: 120 min: 120 min:65.1 ± 8.9 43.4 ± 7.9 293.8 ± 68.9 752.7 ± 129.4 168.5 ± 61.5 603.5 ±97.4 191.8 ± 37.1 180 min: 180 min: 180 min: 180 min:  40.6 ± 17.1 22.8± 9.4 270.5 ± 73.8 212.5 ± 65.1 Small  20 min:  20 min:  20 min:  20min:  20 min:  20 min:  20 min: Movement 767.3 ± 46.7 697.2 ± 36.8 711.6± 65.7 904.3 ± 110.8 589.2 ± 50.4 919.6 ± 96.4 1537.0 ± 97.3 Occurrences  60 min:  60 min:  60 min:  60 min:  60 min:  60 min:  60min: 357.9 ± 47.3 307.8 ± 56.4 563.5 ± 85.0 1218.7 ± 178.9  232.8 ± 33.6 965.2 ± 153.9  992.0 ± 126.7 120 min: 120 min: 120 min: 120 min: 120min: 120 min: 120 min: 256.3 ± 42.9 140.3 ± 22.3  717.4 ± 138.6 1358.1 ±137.1   443.1 ± 100.6 1201.4 ± 160.9 555.0 ± 87.0 180 min: 180 min: 180min: 180 min: 114.4 ± 38.6 103.2 ± 42.0  651.4 ± 141.2  484.5 ± 114.1

Example 142 Rat Behavioral Assay

The objective of this study was to assess the antidepressant effects oftest compounds 124, 125, 126, and 127 in the behavioral despair assay inrats using a modification of a method described by Porsolt R. D., AntonG., Blavet N., Jalfre M., Behavioural despair in rats: a new modelsensitive to antidepressant treatment, Eur. J. Pharmacol., 1978, 41,379-391. The animals were preconditioned in a pretest session, where therats were individually forced to swim inside a vertical plexiglasscylinder containing water maintained at 19-20° C. After 15 minutes inthe water, they were allowed to dry for 15 minutes in a heatedenclosure. Twenty four hours later, the compounds were administeredintraperitoneal to the animals. One hour after administration of thetest compound, animals were put back into the cylinder containing water.The total duration of immobility was measured during the last 4 minutesof a 6 minute test.

Compound Dose (mg/kg) % variation 126 10 99 2.5 45 127 10 67 2.5 25 12410 91 2.5 6 125 10 55 Nomifensine 3 78

The results are expressed as the percentage of variation of the totalduration of immobility calculated from the mean value of thevehicle-treated group (% variation=[(immobility duration ofvehicle-immobility duration of test compound)/(immobility duration ofvehicle)]×100%). Only compounds which exhibit a statisticallysignificant variation >30% are considered effective in this in vivomodel.

Results

Based on the aforementioned criterion for effectiveness, all fourcompounds are effective at a dose of 10 mg/kg; compound 126 is alsoeffective at 2.5 mg/kg.

Example 143 Rat Behavioral Assay

The objective of this study was to assess the antidepressant effects oftest compounds 124, 125, 126, and 127 in the behavioral despair assay inrats according to the methods described by Porsolt R. D., Anton G.,Blavet N., Jalfre M., Behavioural despair in rats: a new model sensitiveto antidepressant treatment, Eur. J. Pharmacol., 1978, 4, 379-391. Theanimals were preconditioned in a pretest session, where the rats wereindividually forced to swim inside a vertical plexiglass cylindercontaining water maintained at 19-20° C. After 15 minutes in the water,they were allowed to dry for 15 minutes in a heated enclosure. Twentyfour hours later, they were replaced in the cylinder and the totalduration of immobility was measured during a 5 minute test (testsession). The test compounds and vehicle were administered as a seriesof 3 intraperitoneal injections 24 h, 5 h and 1 h before the 5 minutetest on the second day.

Compound Dose (mg/kg) % variation 124 2.5 59 2.0 71 1.0 29 125 2.5 12126 1.0 58 126 2.5 76 0.75 47 0.25 9 127 1.0 14 127 3.0 64 2.5 39 2.0 39Imipramine 30 55

The results are expressed as the percentage of variation of the totalduration of immobility calculated from the mean value of thevehicle-treated group (% variation=[(immobility duration ofvehicle-immobility duration of test compound)/(immobility duration ofvehicle)]×100%). Only compounds which exhibit a statisticallysignificant variation >30% are considered effective in this in vivomodel.

Results

Based on the aforementioned criterion for effectiveness, compounds 124,126, and 127 were effective at a dose of 2.5 mg/kg; compound 126 wasalso effective at a dose of 1.0 mg/kg.

Example 144 Determination of the Absolute Stereochemistry of Compound124

The absolute stereochemistry of 124 was determined to be (3S,1′S), byboth asymmetric synthesis and X-Ray crystallography. The method ofenantiospecific preparation of 124 is depicted in Schemes 1 and 2. Thestereocenter at the 3-position of the piperidine was set as the S-isomeraccording to the literature precedent (See Reference 2). The secondstereocenter for the carbinol position was predicted to be the S-isomerby stereoselective reduction of the ketone to give S-epoxide (SeeReference 1). Coupling of the 3S-piperidine with the S-epoxide afforded124. The 1′-stereocenter was confirmed as the S-isomer by itsrelationship to the 3-position stereocenter through X-raycrystallography (See FIG. 1). The measured optical rotation for 124 was[α]_(D)=−263.8° (c=1.5; CHCl₃; 589 nm, 21° C.).

The (3S,1′R) diastereomer, 125, was prepared by coupling the R-epoxidewith the S-piperidine (See Scheme 3). This compound had a unique ¹H-NMRand ¹³C-NMR compared to 124 as expected for a diastereomer. Thiscompound had a measured optical rotation of [α]_(D)=−1545.6° (c=2.26;CHCl₃; 589 nm, 21° C.).

The (3R,1′S) diastereomer, 126, was prepared by coupling the S-epoxidewith the R-piperidine (See Scheme 3). This compound had identical ¹H-NMRand ¹³C-NMR to its enantiomer, 125. This compound had a measured opticalrotation Of [α]_(D)=+1479.1° (c=1.13; CHCl₃; 589 nm, 21° C.).

The (3R,1′R) diastereomer, 127, was prepared by coupling the R-epoxidewith the R-piperidine (See Scheme 3). This compound had identical ¹H-NMRand ¹³C-NMR to its enantiomer, 124. This compound had a measured opticalrotation of [α]_(D)=+258.7° (c=1.50; CHCl₃; 589 nm, 21° C.).

REFERENCES CITED IN EXAMPLE 144

-   1) (a) Corey, E. J.; Bakshi, R. K.; Shibata, S. J. Am. Chem. Soc.    1987, 109, 5551. (1) Corey, E. J.; Bakshi, R. K.; Shibata, S.; Chen,    C.-P.; Singh, V. K. J. Am. Chem. Soc., 1987, 109, 7925. (c)    Corey, E. J.; Shibata, S.; Bakshi, R. K. a Org. Chem., 1988, 53,    2861.-   2) Marnus, P.; Thurston, L. S. J Org. Chem. 1991, 56, 1166.

Example 145 Synthesis of(R)-1-[1-(4-Methoxy-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethoxy-phenoxymethyl)-piperidine

Preparation of(R)-[3-(4-trifluoromethoxy-phenoxymethyl)-piperidine-1-carboxylic AcidTert-Butyl Ester]

To a solution of (R)-3-Methanesulfonyloxymethyl-piperidine-1-carboxylicacid tert-butyl ester (3.522 g, 12 mmol) in CH₃CN (100 mL) at roomtemperature was added 4-(trifluoromethoxy)phenol (1.555 mL, 12 mmol) andCs₂CO₃ (7.820 g, 24 mmol). The mixture was heated at reflux for 20hours. After cooling to room temperature, the mixture was filtered andconcentrated in vacuo. The residue was dissolved in 250 mL EtOAc, andwashed with water (125 mL), saturated Na₂CO₃ (125 mL), and water (125mL), The organic layer was dried over Na₂SO₄, filtered and concentratedin vacuo. The residue was purified by flash column chromatography usinga gradient of 0 to 25% EtOAc in hexane to provide the phenyl ether (3.40g, 81%). ¹H NMR (300 MHz, CDCl₃): δ 7.14 (d, J=9.0 Hz, 2H), 6.88 (d,J=9.0 Hz, 2H), 3.76-3.91 (m, 4H), 2.92 (br s, 2H), 1.99-2.05 (m, 1H),1.87-1.92 (m, 1H), 1.67-1.74 (m, 1H), 1.46 (s, 9H), 1.27-1.40 (m, 21).¹³C NMR (CDCl₃): δ 157.7, 155.2, 142.9, 122.7, 120.0, 115.4, 79.7, 70.6,47.1, 44.4, 36.0, 28.6, 27.5, 24.5.

Preparation of (R)-[3-(4-Trifluoromethoxy-phenoxymethyl)-piperidine]

A solution of the phenyl ether (3.40 g, 9.60 mmol) in CH₂Cl₂ (30 mL) at0° C. was treated with TFA (30 mL). The reaction mixture was allowed towarm up to 25° C., and stirred for 1 hour. The solvent was removedNa₂SO₄. The residue was dissolved in CH₂Cl₂ (50 mL), washed withsaturated NaHCO₃ (50 mL) and brine (50 mL). The organic layer was driedover Na₂SO₄, filtered and concentrated in vacuo to yield the secondaryamine (2.31 g, 88%). LRMS m/z 276 (M⁺, C₁₃H₁₆F₃NO₂ ⁺, requires 276).

Preparation of(R)-[1-(Methoxy-phenyl)cyclobutyl]-[3-(4-trifluoromethoxy-phenoxymethyl0-piperidin-1-yl]-methanone

To a solution of the secondary amine (1.0 g, 3.63 mmol),1-(4-methoxy-phenyl)-cyclobutanecarboxylic acid (0.824 g, 4.0 mmol) andHOBt (0.612 g, 4.0 mmol) in DMF (8.0 mL) was added1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide methiodide (1.19 g, 4.0mmol). The mixture was stirred at room temperature for 20 hours. Then,the reaction mixture was poured into water (30 mL), extracted with EtOAc(3×30 mL). The organic layer was washed with saturated NaHCO₃ (30 mL),brine (30 mL), and brine (30 mL), dried over Na₂SO₄, and concentrated invacuo. The residue was purified by flash column chromatography using agradient of 0 to 25% EtOAc/hexane to provide the corresponding amide(1.40 g, 83%). ¹H NMR (CDCl₃): δ 7.33 (d, J=8.7 Hz, 2H), 7.15 (d, J=8.7Hz, 2H), 6.76-6.91 (m, 4H), 4.50 (br s, 1H), 3.80 (s, 3H), 3.28-3.65 (m,3H), 2.30-2.94 (m, 6H), 1.73-2.06 (m, 4H), 1.25-1.45 (m, 3H). ¹³C NMR(CDCl₃): δ 174.6, 158.3, 157.5, 142.9, 135.9, 126.4, 122.7, 120.0,115.4, 115.2, 114.3, 70.5, 55.4, 52.0, 49.0, 46.3, 45.6, 43.2, 36.0,33.1, 32.4, 27.4, 24.6, 24.0, 15.6.

Preparation of(R)-1-[1-(4-Methoxy-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethoxy-phenoxymethyl)-piperidine

To a solution of the amide (0.70 g, 1.51 mmol) in THF (20 mL) was addedLiAlH₄ (0.058 g, 1.51 mmol) at −70° C. After addition, the reactionmixture was heated at reflux for 3 hours. Then, the mixture was cooledto 0° C., and quenched with 2 N NaOH (0.3 mL) and water (0.3 mL). Themixture was filtered and concentrated in vacuo. The residue was purifiedby flash column chromatography, eluting with CH₂Cl₂/MeOH to provide freeamine 170 (0.58 g, 86%, >99.9% ee). ¹H NMR (300 MHz, CDCl₃): δ 7.12-7.19(m, 4H), 6.89 (d, J=2.6 Hz, 2H), 6.85 (d, J=2.6 Hz, 2H), 3.82 (s, 3H),3.65-3.78 (m, 2H), 2.63 (AB quartet, J=13.5 Hz, 2H), 1.11-2.54 (m, 15H).¹³C NMR (CDCl₃): δ 158.0, 157.5, 142.7, 142.2, 127.3, 122.6, 120.0,115.4, 113.3, 71.3, 69.4, 59.1, 56.6, 55.4, 47.0, 36.3, 32.0, 31.9,27.0, 24.9, 16.3. LRMS m/z 450 (M⁺, C₂₅H₃₀F₃NO₃ ⁺, requires 450).

Example 146 Synthesis of(S)-1-[1-(4-Methoxy-phenyl)-cyclobutylmethyl]-3-(4-trifluoromethoxy-phenoxymethyl)-piperidine

Compound 171 was prepared using the procedure outlined in Example 145,starting with the (S)-3-methanesulfonyloxymethyl-piperidine-1-carboxylicacid tert-butyl ester. The chiral purity of 171 was determined to be98.4% ee using chiral HPLC analysis.

Example 147 Synthesis of[2-{3-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-cyclohexyl}-2-(4-trifluoromethyl-phenoxy)-ethyl]-dimethyl-amine

The epoxide (10.0 g) was dissolved in 200 mL of 2.0 M dimethylamine inTHF in a sealed tube. The mixture was stirred at 60° C. for 48 hours.After removal of the solvent, the residual was purified on columnchromatography (silica gel, EtOAc to EtOAc/MeOH, 1:1). 5.56 g of aminoalcohol was obtained (LRMS 273).

The amino alcohol (8.95 g, 32.9 mmol) was dissolved in 80 mL of CH₂Cl₂and cooled down at 0° C. Then N,N-diisopropylethylamine (11.46 mL, 2eq.) and methanesulfonyl chloride (3.06 mL, 1.2 eq.) were added. Thereaction mixture was stirred at r.t. for 6 hours. To the mixture wasadded 20 mL of water and the aqueous layer was extracted with EtOAc(3×100 mL). The organic layer was washed with brine and dried overNa₂SO₄. After filtration and evaporation, the crude residual (10.5 g)was dissolved in 90 mL of CH₃CN. To the solution the potassium carbonate(20.7 g, 5 eq.) and α,α,α,-trifluoro-p-cresol (9.72 g, 2 eq.) were addedand the mixture was stirred at 60° C. overnight. The mixture wasquenched with water (50 mL), extracted with EtOAc (2×60 mL). Thecombined organic phase was washed with brine (2×50 mL) and dried overNa₂SO₄. After filtration and removal of the solvent, silica gel flashcolumn chromatography gave the diastereomeric phenyl ethers.

To the diastereomer shown (25 mg, 0.06 mmol) in CH₂Cl₂ (1.0 mL),trifluoroacetic acid (1.0 mL) was added at 0° C. After completion ofaddition the reaction mixture continued sting at r.t. for 20 minutes.The solvent was removed and the residual was dried in vacuo for 2 hours.The resulting oil was dissolved in 1.0 mL of DMF. To this solution1-(4-chlorophenyl)-1-cyclobutanecarboxylic acid (15.2 mg, 1.2 eq.),1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide methadone (21.4, 1.2eq.) and HOBt were added. The mixture was stirred at r.t. overnight. Thereaction was quenched with 20 mL of EtOAc and 5 mL of 10% aqueous NaOH.The organic layer was washed with 5 mL of brine and dried over Na₂SO₄.The preparative TLC (silica gel, EtOAc/Hexane, 2:1) gave (+/−)-173 (27mg, yield 89%). LRMS 509.

To a solution of (+/−)-173 (25 mg, 0.05 mmol) in 3 mL of dry THF wasadded LiAlH₄ (4 mg) at r.t. The mixture was refluxed for 2 hour. Thereaction mixture was quenched with 5 mL of water and extracted withEtOAc (3×10 mL). The combined organic layer was washed with brine (2×5mL) and dried over Na₂SO₄. The preparative TLC (silica gel, EtOAc) gave(+/−>174 (13 mg, yield 51%). LRMS 495.

Employing the same procedures, compound the diastereomer shown (25 mg)was converted into compound (+/−)-175 (10 mg, LRMS 495).

Example 148 Synthesis of1-(2,3-Dihydro-benzo[1,4]dioxin-2-ylmethyl)-3-(4-trifluoromethyl-phenoxymethyl)-piperidine

Racemic 3-(4-Trifluoromethyl-phenoxymethyl)-piperidine (100 mg, 0.39mmol), R-2,3-Dihydro-benzo[1,4]dioxine-2-carboxylic acid (73 mg, 0.41mmol), the amide coupling agent BrOP (225 mg, 0.58 mmol), anddiisopropylethylamine (150 mg, 1.16 mmol) were dissolved in 2 mL ofanhydrous dichloromethane. Concomitantly, racemic3-(4-Trifluoromethyl-phenoxymethyl)-piperidine (100 mg, 0.39 mmol),S-2,3-Dihydro-benzo[1,4]dioxine-2-carboxylic acid (73 mg, 0.41 mmol),the amide coupling agent BrOP (225 mg, 0.58 mmol), anddiisopropylethylamine (150 mg, 1.16 mmol) were dissolved in 2 mL ofanhydrous dichloromethane. Each mixture was kept at room temperatureovernight, diluted with 10 mL of water, and 10 mL of ether. Extractiveworkup gave in each case, after concentration of the organic layers invacuo and chromatography on silica gel using a EtOAc-hexane gradientcolumn, the desired amide intermediate in 75-88% yield.

Each intermediate amide mixture was dissolved in 5 mL of THF at 0° C.and excess lithium aluminum hydride (100 mg, ca. 7 equivalents) wasadded. The solutions were heated to reflux for 5 minutes, cooled to 0°C., and quenched by dropwise addition of 0.5 mL 1M NaOH. Additional THF(10 mL) was added to each, and the suspensions were stirred at roomtemperature for 30 minutes and then filtered though a plug of sodiumsulfate. The solutions thus obtained were concentrated in vacuo and theresidues were purified by preparative HPLC using a Chiralpak AD™ columnfrom Chiral Technologies, Inc., eluting with an 99:1 mixture of hexaneand isopropyl alcohol containing ca. 0.1% diethylamine. Each mixture ofdiastereomers was separated in this fashion, and the pairs of amineswere obtained in 50-65% total yield from each corresponding amidemixture (176, 177, 178, and 179). Data for each diastereomer: MS 408(M⁺+1). ¹H and ¹³C (DEPT) NMR data for each isomer was consistent withthe assigned structure. Particularly diagnostic in distinguishing thediastereomers by NMR was the observance of a pair of doublets in the ¹Hspectrum that appear at 6=3.09 ppm (J=10 Hz) and 6=2.80 ppm (J=10 Hz)for one diastereomer and 6=2.98 ppm (J=7 Hz) and 6=2.91 ppm (J=7 Hz) forthe other diastereomer.

Example 149 Synthesis of1-(4-Chloro-phenoxy)-3-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-propan-2-ol

2-(4-Chloro-phenoxymethyl)-oxirane (50 mg) and of3-(4-Trifluoromethyl-phenoxymethyl)-piperidine (70 mg, 1.0 equivalent)in 4 mL of methanol were heated in a sealed tube at 100° C. for 16hours, cooled to room temperature, transferred to a round-bottomedflask, and concentrated in vacuo. The crude residue was purified byflash chromatography on silica gel using as eluent a gradient of ethylacetate in hexane containing 1% ammonium hydroxide. 95 mg of1-(4-Chloro-phenoxy)-3-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-propan-2-olwas obtained (79%). Data for this mixture of amino alcoholdiastereomers: MS 444 (M+1).

Example 150 Synthesis of3-{2-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethyl}-1H-indole

3-(4-Trifluoromethyl-phenoxymethyl)-piperidine (50 mg, 0.19 mmol),(1H-Indol-3-yl)-acetic acid (73 mg, 0.21 mmol), the amide coupling agentBrOP (123 mg, 0.32 mmol), and diisopropylethylamine (75 mg, 0.58 mmol)were dissolved in 2 mL of anhydrous dichloromethane. The mixture waskept at room temperature overnight, diluted with 10 mL of water, and 10mL of ether. Extractive workup gave in each case, after concentration ofthe organic layers in vacuo and chromatography on silica gel using aEtOAc-hexane gradient column, the desired amide intermediate (71 mg,90%). This amide was dissolved in 5 mL of THF at 0° C. and excesslithium aluminum hydride (50 mg, ca. 8 equivalents) was added. Thesolution was heated to reflux for 5 minutes, cooled to 0° C., andquenched by dropwise addition of 0.5 mL 1M NaOH. Additional THF (10 mL)was added, and the suspension was stirred at room temperature for 30minutes and then filtered though a plug of sodium sulfate. The solutionthus obtained was concentrated in vacuo and the residue was purified byflash chromatography on silica gel using as eluent a gradient of ethylacetate in hexane containing 1% ammonium hydroxide. 53 mg of3-{2-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethyl}-1H-indole,180, was obtained (77%). Data for this compound: MS 403 (M⁺+1).

Example 151 Synthesis of1-(2-Biphenyl-4-yl-ethyl)-3-(4-trifluoromethyl-phenoxymethyl)-piperidine

Following exactly the same two step procedure used in the previousexample for the preparation of3-{2-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethyl}-1H-indolebut using biphenyl-4-yl-acetic acid in place of (1H-Indol-3-yl)-aceticacid,1-(2-Biphenyl-4-yl-ethyl)-3-(4-trifluoromethyl-phenoxymethyl)-piperidinewas obtained in 54% overall yield from 0.19 mmol of3-(4-Trifluoromethyl-phenoxymethyl)-piperidine, 181. Data for thiscompound: MS 440 (M⁺+1).

Example 152 Synthesis of(S)-[2-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-1-phenyl-ethanol]

Preparation of (S)-[3-(2-Oxo-2-phenyl-ethyl)-piperidine-1-carboxylicAcid Benzyl Ester]

To a solution of (S)-3-styryl-piperidine-1-carboxylic acid benzyl ester(411 mg, 1.28 mmol, in 5 mL of THF) was added 3.86 mL of a 0.5 M THFsolution of 9-borabicyclononane. The solution was brought to reflux andthen cooled to room temperature after 3 hours. 2 mL of 30% hydrogenperoxide solution and 2 mL of 3M NaOH were added dropwise. The solutionwas stirred for 30 minutes, diluted with water (20 mL) and ether (30mL). Extractive workup gave, after concentration of the organic layersin vacuo and chromatography on silica gel using as eluent anEtOAc-hexane gradient, the desired alcohol intermediate (69 mg, 85%; MS340=M⁺+1). This alcohol (350 mg, 1.03 mmol) was dissolved in 5 mL ofmethylene chloride and treated with 1.1 equivalents of the Dess-Martinperiodinane. After 2 hours at room temperature, 1 mL of isopropanol wasadded, and then after 5 minutes 5 mL of 1M NaOH was added. After 20minutes, to the mixture was added 30 mL of ether and 30 mL of water.Extractive workup gave, after concentration of the organic layers invacuo and chromatography on silica gel using as eluent an EtOAc-hexanegradient, the desired ketone intermediate(S)-3-(2-Oxo-2-phenyl-ethyl)-piperidine-1-carboxylic acid benzyl ester(337 mg, 97%; MS 338=M⁺+1).

Preparation of (S)-[1-phenyl-2-piperidin-3-yl-ethanone]

(S)-[3-(2-Oxo-2-phenyl-ethyl)-piperidine-1-carboxylic acid benzyl ester](266 mg, 0.79 mmol) was dissolved in 2 mL of methanol in a small vesselfor pressurized hydrogenation. 200 mg of 5% palladium on carbon wasadded. The vessel was charged with 50 psi of hydrogen and shaken for twohours. The vessel was evacuated, filtered, and the residue wasconcentrated in vacuo to yield 155 mg of the desired product(S)-[1-Phenyl-2-piperidin-3-yl-ethanone] (97%; MS 204=M⁺+1).

Preparation of(S)-[2-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-1-phenyl-ethanol]

(S)-[1-Phenyl-2-piperidin-3-yl-ethanone] (142 mg, 0.70 mmol) and1-(4-Chloro-phenyl)-cyclobutanecarboxylic acid (148 mg, 0.70 mmol) weredissolved in 5 mL of methylene chloride and treated with the amidecoupling agent BrOP (1.05 mmol), and diisopropylethylamine (2.1 mmol).The mixture was kept at room temperature overnight, diluted with 20 mLof water, and 50 mL of ether. Extractive workup gave, afterconcentration of the organic layers in vacuo and chromatography onsilica gel using a EtOAc-hexane gradient column, the desired amideintermediate2-{1-[1-(4-Chloro-phenyl)-cyclobutanecarbonyl]-piperidin-3-yl)}-1-phenyl-ethanone(254 mg, 92%). This amide was dissolved in 10 mL of THF at 0° C. andexcess lithium aluminum hydride (245 mg) was added. The solution washeated to reflux for 5 minutes, cooled to 0° C., and quenched bydropwise addition of 1.5 mL 1M NaOH. Additional THF (10 mL) was added,and the suspension was stirred at room temperature for 30 minutes andthen filtered though a plug of sodium sulfate. The solution thusobtained was concentrated in vacuo and the residue was purified by flashchromatography on silica gel using as eluent a gradient of ethyl acetatein hexane containing 1% ammonium hydroxide. 205 mg of(S)-[2-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-1-phenyl-ethanol],182, was obtained (83%). ¹H and ¹³C NMR data was consistent with theassigned structure. Data for this compound as a mixture ofdiastereomers: MS 384 (++1).

Example 153 Synthesis of(S)-[2-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-1-phenyl-ethanone]

(S)-[2-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-1-phenyl-ethanol](29 mg, 0.075 mmol) was dissolved in 2 mL of methylene chloride andtreated with 1.1 equivalents of the Dess-Martin periodinane. After 2hours at room temperature, 0.5 mL of isopropanol was added, and thenafter 5 minutes 1 mL of 1M NaOH was added. After 20 minutes, to themixture was added 20 mL of ether and 20 mL of water. Extractive workupgave, after concentration of the organic layers in vacuo andchromatography on silica gel using as eluent an EtOAc-hexane gradientwith 1% ammonium hydroxide, the desired product(S)-[2-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-1-phenyl-ethanone],183, (15 mg, 53%). Data for this ketone: MS 382 (M⁺+1).

Example 154 Synthesis of(S)-[1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(2-methoxy-2-phenyl-ethyl)-piperidine]

(S)-[2-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-1-phenyl-ethanol](38 mg, 0.100 mmol) was dissolved in 1 mL of THF and cooled to 0° C.Potassium tert-butoxide was added (112 mg, 1.0 mmol) followed by methyliodide in large excess (ca. 0.5 mL). After 30 minutes at 0° C., water (5mL) and ether (5 mL) were added. Extractive workup gave, afterconcentration of the organic layers in vacuo and chromatography onsilica gel using as eluent an EtOAc-hexane gradient with 1% ammoniumhydroxide, the desired product1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(2-methoxy-2-phenyl-ethyl)-piperidine,184, (28 mg, 70%). Data for this compound: MS 398 (M⁺+1).

Example 155 Synthesis of(S)-[1-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-2-phenyl-propan-2-ol]

(S)-[2-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-1-phenyl-ethanone](10 mg, 0.027 mmol) was dissolved in 1 mL of anhydrous THF and cooled to0° C. Methyl magnesium bromide solution (0.27 mL of a 1M solution inTHF, 0.27 mmol) was added via syringe. After 30 minutes the reaction wasquenched by adding 0.5 mL of 1M NaHSO₄ solution. Water (4 mL) and ether(10 mL) were added. Extractive workup gave, after concentration of theorganic layers in vacuo and chromatography on silica gel using as eluentan EtOAc-hexane gradient with 1% ammonium hydroxide, the desired product(S)-[1-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-2-phenyl-propan-2-ol],185, (8 mg, 75%). Data for this compound: MS 398 (M⁺+1).

Example 156 Synthesis of(S)-1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(2-phenyl-propyl)-piperidine

(S)-2-{1-[1-(4-Chloro-phenyl)-cyclobutanecarbonyl]-piperidin-3-yl}-1-phenyl-ethanone(12.7 mg, 0.032 mmol) in 1 mL of THF was treated with 5 equivalents ofmethylene Wittig reagent at room temperature. The crude mixture wasconcentrated in vacuo, dissolved in a minimal amount of methylenechloride, and then this product mixture purified by flash columnchromatography on silica gel using a gradient elution with anEtOAc-hexane mixture. The desired olefin was obtained (10.4 mg, 82%) andwas immediately dissolved in 1 mL of methanol in a small vessel forpressurized hydrogenation. 20 mg of 10% palladium on carbon was added.The vessel was charged with 50 psi of hydrogen and shaken for 6 hours.The vessel was evacuated, filtered, and the residue was concentrated invacuo to yield 10.3 mg of the desired product (99%). This unpurifiedproduct was dissolved in 1 mL of THF at 0° C. and excess lithiumaluminum hydride (0.30 mL of a 1M solution in ether, ca. 10 equiv.) wasadded. The solution was heated to reflux for 5 minutes, cooled to 0° C.,and quenched by dropwise addition of 0.5 mL 1M NaOH. Additional THF (2mL) was added, and the suspension was stirred at room temperature for 30minutes and then filtered though a plug of sodium sulfate. The solutionthus obtained was concentrated in vacuo and the residue was purified byflash chromatography on silica gel using as eluent a gradient of ethylacetate in hexane containing 1% ammonium hydroxide.(S)-1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-3-(2-phenyl-propyl)-piperidine,186, was obtained (7.2 mg, 72%). Data for this compound: MS 382 (M⁺+1).

Example 157 Synthesis of1-(4-Chloro-phenyl)-4-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-ylmethyl]-cyclohexanol

3-(4-Trifluoromethyl-phenoxymethyl)-piperidine (122 mg, 0.47 mmol),4-Oxo-cyclohexanecarboxylic acid (67 mg, 0.47 mmol) the amide couplingagent BrOP (274 mg, 0.71 mmol), and diisopropylethylamine (183 mg, 1.41mmol) were dissolved in 2 mL of anhydrous dichloromethane. The mixturewas kept at room temperature overnight, diluted with 10 mL of water, and10 mL of ether. Extractive workup gave, after concentration of theorganic layers in vacuo and chromatography on silica gel using aEtOAc-hexane gradient column, the desired amide intermediate,4-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidine-1-carbonyl]-cyclohexanone(136 mg, 75%; MS 384=M⁺+1).

A portion of this ketone (105 mg, 0.27 mmol) amide mixture was dissolvedin 2 mL of THF at 0° C. and p-C₁-phenyl magnesium bromide (0.68 mL of a1M solution, 2.5 equivalents) was added via syringe. After 30 minutes, 2mL of 1M NaHSO₄ solution was added, followed by 15 mL of water and 15 mLof ether. Extractive workup gave, after concentration of the organiclayers in vacuo and chromatography on silica gel using a EtOAc-hexanegradient column, the desired Grignard adduct[4-(4-Chloro-phenyl)-4-hydroxy-cyclohexyl]-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-methanone(88 mg, 65%; MS 478=M+1 minus water). A portion of this material (41 mg,0.083 mmol) was dissolved in 2 mL of THF and excess lithium aluminumhydride (31 mg, ca 10 equivalents) was added. The solution was heated toreflux for 5 minutes, cooled to 0° C., and quenched by dropwise additionof 1 mL 1M NaOH. Additional THF (10 mL) was added, and the suspensionwas stirred at room temperature for 30 minutes and then filtered thougha plug of sodium sulfate. The solution thus obtained was concentrated invacuo and the residue was purified by flash chromatography on silica gelusing as eluent a gradient of ethyl acetate in hexane containing 1%ammonium hydroxide.1-(4-Chloro-phenyl)-4-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-ylmethyl]-cyclohexanol(187) was obtained (28 mg, 70%). MS 482 (M⁺+1), 464 (M⁺+1 minus water).

Example 158 Synthesis of4-[3-(4-Trifluoromethyl-phenoxymethyl)piperidin-1-ylmethyl]-cyclohexanol

4-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidine-1-carbonyl]-cyclohexanone(27 mg, 0.070 mmol) was dissolved in 1 mL of THF and excess lithiumaluminum hydride (27 mg, ca. 10 equivalents) was added. The solution washeated to reflux for 5 minutes, cooled to 0° C., and quenched bydropwise addition of 0.5 mL 1M NaOH. Additional THF (10 mL) was added,and the suspension was stirred at room temperature for 30 minutes andthen filtered though a plug of sodium sulfate. The solution thusobtained was concentrated in vacuo and the residue was purified by flashchromatography on silica gel using as eluent a gradient of ethyl acetatein hexane containing 1% ammonium hydroxide.4-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidin-1-ylmethyl]-cyclohexanol,188, (16 mg, 61%) was obtained. Data for this compound: MS 372 (M⁺+1).

Example 159 Synthesis of3-(3,5-Bis-trifluoromethyl-benzyloxy)-1-[1-(4-chloro-phenyl)-cyclobutylmethyl]-piperidine

Following the procedure described in Example 72,1-Bromomethyl-3,5-bis-trifluoromethyl-benzene was used to alkylate 56 mg(0.19 mmol) of[1-(4-Chloro-phenyl)-cyclobutyl]-(3-hydroxy-piperidin-1-yl)-methanone,forming the ether adduct[3-(3,5-Bis-trifluoromethyl-benzyloxy)-piperidin-1-yl]-[1-(4-chloro-phenyl)-cyclobutyl]-methanone(39 mg, 39%). A portion of this compound (22 mg, 0.043 mmol) wasdissolved in 1 mL of THF and excess lithium aluminum hydride (16 mg, ca.10 equivalents) was added. The solution was heated to reflux for 5minutes, cooled to 0° C., and quenched by dropwise addition of 0.5 mL 1MNaOH. Additional THF (10 mL) was added, the suspension was stirred atroom temperature for 30 minutes, and then was filtered though a plug ofsodium sulfate. The solution thus obtained was concentrated in vacuo andthe residue was purified by flash chromatography on silica gel using aseluent a gradient of ethyl acetate in hexane containing 1% ammoniumhydroxide.3-(3,5-Bis-trifluoromethyl-benzyloxy)₁₋₁-[1-(4-chloro-phenyl)-cyclobutylmethyl]-piperidine,189, was obtained (12 mg, 56%). Data for this compound: MS 506 (M⁺+1).

Example 160 Synthesis of2-Cyclohexyl-2-hydroxy-2-phenyl-1-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone

A solution of 3-(4-trifluoromethyl-phenoxymethyl)-piperidine (0.220mmol, 57 mg) and cyclohexyl-hydroxy-phenyl-acetic acid (1.2 equiv, 0.264mmol, 62 mg) in CH₂Cl₂ (1 mL) was treated with BrOP (1.5 equiv, 0.330mmol, 314 mg) and iPr₂NEt (3.0 equiv, 0.660 mmol, 115 μL) at 0° C. Thereaction mixture stirred for 12 h while warming to rt. The reactionmixture was quenched with 10% HCl (10 mL) and then extracted with EtOAc(2×15 mL). The combined organics were washed with NaHCO_(3(sat)) anddried with NaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed invacuo and chromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm, 5:1hexane-ethyl acetate) provided 190 (45 mg, 105 mg theoretical, 43%) as acolorless oil: R_(f) 0.45 (SiO₂, 5:1 hexane-ethyl acetate); LRMS m/z 476(M⁺+1, C₂₇H₃₂F₃NO₃ requires 476).

Example 161 Synthesis of1-Cyclohexyl-1-phenyl-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A solution of 190 (0.095 mmol, 45 mg) in THF (500 μL) at 0° C. wastreated with LiAlH₄ (5.0 equiv, 0.475 mmol, 18 mg) under Ar. Thereaction mixture stirred for 12 h and returned to 25° C. The reactionmixture was then cooled to 0° C., quenched with 10% aqueous NaOH andextracted with 3×EtOAc (25 mL). The organics were dried withNaCl_((sat)) and Na₂SO_(4(s)). The reaction mixture was purified bychromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm, 4:1 hexane-ethyl acetate)which provided 191 (34 mg, 44 mg theoretical, 77%) as a colorless oil:R_(f) 0.38 (SiO₂, 4:1 hexane-ethyl acetate); LRMS m/z 462 (M⁺+1,C₂₇H₃₄F₃NO₂ requires 462).

Example 162 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-{3-[2-(4-trifluoromethyl-phenyl)-ethyl]-piperidin-1-yl}-ethanol,Separation of all diastereomers

A solution of 155 (0.293 mmol, 136 mg) in CH₃OH (2 mL) was treated withNaBH (3.0 equiv, 0.879 mmol, 33 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (10 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_(4(sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 35 g cartridge, 2:1 hexane-ethyl acetate) provided amixture of diastereomers (192, 193, 194, and 195) (120 mg, 137 mgtheoretical, 88%) as a colorless oil: R_(f) 0.35 (SiO₂, 2:1 hexane-ethylacetate); LRMS m/z 467 (M⁺+1, C₂₆H₃₁ClF₃NO requires 467). The fourdiastereomers were separated on a Chiralpak AD column by utilizing thefollowing procedure. The mixture was dissolved in 90:10 hexane (0.1%diethylamine) and isopropanol at a concentration of 90 mg/mL. 192 (firstpeak) and 193 (fourth peak) were separated by using a 99% hexane (0.1%diethylamine) and 1% isopropanol solvent system. The middle peak wascollected and concentrated in vacuo and then separated using 95% hexane(0.1% diethylamine) and 5% isopropanol to provide 194 (second peak) and195 (third peak).

Example 163 Synthesis of1-[2-(4-Chloro-phenyl)-2-methoxy-ethyl]-3-(R)-phenethyl-piperidine

A solution of 233 (0.378 mmol, 130 mg) in THF (1 mL) and CH₃I (1 mL) wastreated with tBuOK (5.0 equiv, 1.89 mmol, 212 mg) at 25° C. The reactionmixture stirred for 10 min. The reaction mixture was quenched with pH 7phosphate buffer (10 mL) and extracted with EtOAc (2×10 mL). Thecombined organics were dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and the residue was purified bychromatography (PTLC, SiO₂, cm×20 cm, 1 mm, 3:1 hexane-acetone) whichprovided 196 (100 mg, 135 mg theoretical, 74%) as a colorless oil: R_(f)0.48 (SiO₂, 3:1 hexane-acetone); LRMS m/z 359 (M⁺+1, C₂₂H₂₈ClNO requires359).

Example 164 Synthesis of2-(4-Chloro-phenyl)-1-(3(R)-phenethyl-piperidin-1-yl)-propan-2-ol

A solution of piperidine-ketone (0.693 mmol, 237 mg) in THF (1 mL) wasadded to a solution of CH₃MgCl (3.0 M in THF) (5.0 equiv, 3.47 mmol,1.16 ml) in THF (1 mL) at 0° C. The reaction mixture stirred for 1 h.The reaction mixture was quenched with 10% NaOH (10 mL) and extractedwith EtOAc (2×10 mL). The combined organics were dried with NaCl_((sat))and Na₂SO_(4(s)). The solvents were removed in vacuo and the residue waspurified by chromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm, 3:1hexane-acetone) which provided 197 (220 mg, 248 mg theoretical, 89%) asa colorless oil: R_(f) 0.44 (SiO₂, 3:1 hexane-acetone); LRMS m/z 359(M⁺+1, C₂₂H₂₈ClNO requires 359).

Example 165 Synthesis of3-[2-(4-Trifluoromethoxy-phenyl)-vinyl]-piperidine-1-carboxylic AcidBenzyl Ester

A solution of the Wittig salt (1.5 equiv, 12.02 mmol, 6.22 g) in THF (40mL) was treated with nBuLi (1.5 equiv, 2.5M, 12.02 mmol, 4.8 mL) at −78°C. The solution was warmed to 0° C. for 30 min and then cooled again to−78° C. A solution of piperidine-3-carbaldehyde (8.01 mmol, 1.98 g) inTHF (10 mL) was added to the above reaction mixture at −78° C. Thereaction stirred for 12 h. The reaction mixture was quenched with 10%HCl (20 mL) and then extracted with EtOAc (2×50 mL). The combinedorganics were dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (Isco Combi-Flash, 110 gcartridge, 19:1 hexane-ethyl acetate) provided 198 (1.79 g, 3.25 gtheoretical, 55%) as a colorless oil: R_(f) 0.42 (SiO₂, 6:1hexane-EtOAc); LRMS m/z 406 (M⁺+1, C₂₂H₂₂F₃NO₃ requires 406).

Example 166 Synthesis of3-[2-(4-Trifluoromethoxy-phenyl)-ethyl]-piperidine

A solution of 198 (4.42 mmol, 1.79 g) in CH₃OH (60 mL) was treated 30%Pd—C (500 mg) and H₂ (Parr Hydrogenator, starting 65 psi). The reactionwas shaken for 4 h. The reaction mixture was filtered through celite,and the solvents were removed in vacuo to provide 199 (1.21 g, 1.21 gtheoretical, quantitative) as a colorless oil: LRMS m/z 274 (M⁺+1,C₁₄H₁₈F₃NO requires 274).

Example 167 Synthesis of1-(4-Chloro-phenyl)-2-{3-[2-(4-trifluoromethoxy-phenyl)-ethyl]-piperidin-1-yl}-ethanone

A solution of 199 (3.66 mmol, 1.00 g), 2-bromo-4′-chloroacetophenone(1.0 equiv, 3.66 mmol, 855 mg) and KF (50% wt on celite) (8.0 equiv,29.28 mol, 1.70 g) in CH₃CN (12 mL) was stirred for 12 h at 25° C. Thereaction mixture was filtered, and the solvents were removed in vacuo.Chromatography (Isco Combi-Flash, 35 g cartridge, 6:1 hexane-acetone)provided 200 (400 mg, 1.57 g theoretical, 25%) as a colorless oil: R_(f)0.49 (SiO₂, 6:1 hexane-acetone); LRMS m/z 429 (M⁺+1, C₂₂H₂₃ClF₃NO₂requires 429).

Example 168 Synthesis of1-(4-Chloro-phenyl)-2-{3-[2-(4-trifluoromethoxy-phenyl)-ethyl]-piperidin-1-yl}-ethanol

A solution of 200 (0.829 mmol, 353 mg) in CH₃OH (4 mL) was treated withNaBH (1.5 equiv, 1.24 mmol, 47 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (5 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 35 g cartridge, 2:3 hexane-ethyl acetate) provided201 (200 mg, 355 mg theoretical, 56%) as a colorless oil: R_(f) 0.36(SiO₂, 2:3 hexane-ethyl acetate); LRMS m/z 429 (M⁺+1, C₂₂H₂₅ClF₃NO₂requires 429).

Example 169 Synthesis of1-(4-Trifluoromethoxy-phenyl)-cyclobutanecarbonitrile

A solution of (4-trifluoromethoxy)-acetophenyl nitrile (9.94 mmol, 2.00g), dibromopropane (1.1 equiv, 10.93 mmol, 1.1 mL) and NaH (60% wt inmineral oil) (2.5 equiv, 24.85 mol, 1.00 g) in DMSO (35 mL) was stirredfor 12 h. The reaction mixture was quenched with pH 7 phosphate buffer(50 mL) and extracted with EtOAc (2×50 mL). The combined organics weredried with NaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed invacuo and chromatography (Isco Combi-Flash, 110 g cartridge, 6.5:1hexane-ethyl acetate) provided 202 (1.77 g, 2.42 g theoretical, 73%) asa colorless oil.

Example 170 Synthesis of1-[1-(4-Trifluoromethoxy-phenyl)-cyclobutyl]-ethanone

A solution of 202 (4.11 mmol, 1.00 g) in toluene (5 mL) was treated withCH₃MgBr (3.0 M in ether) (3.0 equiv, 12.33 mol, 4.2 mL). The reactionwas stirred for 12 h at 95° C. The reaction mixture was quenched with 6M HCl and stirred for 1 h at 95° C. The reaction mixture was extractedwith EtOAc (2×50 mL). The combined organics were dried withNaCl_(4(sat)) and Na₂SO_(4(s)). The solvents were removed in vacuo andchromatography (Isco Combi-Flash, 110 g cartridge, 30:1 hexane-ethylacetate) provided 203 (0.939 g, 1.06 g theoretical, 89%) as a colorlessoil.

Example 171 Synthesis of2-Bromo-1-[1-(4-trifluoromethoxy-phenyl)-cyclobutyl-ethanone

A solution of 203 (1.94 mmol, 500 mg) in CH₃OH (5 mL) was cooled to 0°C. and treated with AcOH (30% HBr) (50 μL). Br₂ (1.94 mmol, 100 μL) wasthen added slowly at 0° C. The reaction mixture was stirred for 12 h at5° C. The reaction mixture was quenched with H₂O (40 mL) and extractedwith EtOAc (2×50 mL). The combined organics were dried with NaCl_((sat))and Na₂SO_(4(s)). The solvents were removed in vacuo to give 204 (500mg, 654 mg theoretical, 76%) as a colorless oil.

Example 172 Synthesis of1-[1-(4-Trifluoromethoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone

A solution of 3-(4-trifluoromethyl-phenoxymethyl)-piperidine (0.359mmol, 93 mg), 204 (1.0 equiv, 0.430 mmol, 145 mg) and KF (50% wt oncelite) (7.0 equiv, 2.51 mol, 292 mg) in CH₃CN (2 mL) was stirred for 12h at 25° C. The reaction mixture was filtered, and the solvents wereremoved in vacuo. Chromatography (Isco Combi-Flash, 10 g cartridge, 2:1hexane-ethyl acetate) provided 205 (126 mg, 185 mg theoretical, 68%) asa colorless oil: R_(f) 0.34 (SiO₂, 2:1 hexane-ethyl acetate); LRMS m/z516 (M⁺+1, C₂₆H₂₇F₆NO₃ requires 516).

Example 173 Synthesis of1-[1-(4-Trifluoromethoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A solution of 205 (0.244 mmol, 126 mg) in CH₃OH (1 mL) was treated withNaBH₄ (1.5 equiv, 0.366 mmol, 14 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (10 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 35 g cartridge, 2:1 hexane-ethyl acetate) provided206 (107 mg, 126 mg theoretical, 85%) as a colorless oil: R_(f) 0.33(SiO₂, 2:1 hexane-ethyl acetate); LRMS m/z 518 (M⁺+1, C₂₆H₂₉F₆NO₃requires 518).

Example 174 Synthesis of3-(4-Trifluoromethoxy-phenoxymethyl)-piperidine-1-carboxylic Acid BenzylEster

A solution of 3-iodomethylpiperidine-1-carboxylic acid benzyl ester(2.78 mmol, 1.00 g), 4-trifluoromethoxy-phenol (1.1 equiv, 3.06 mmol,545 mg) and Cs₂CO₃ (3.0 equiv, 3.06 mmol, 2.72 g) in CH₃CN (10 mL) washeated to 65° C. The solution was stirred for 12 h. The reaction mixturewas quenched with H₂O (20 mL) and then extracted with EtOAc (2×50 mL).The combined organics were dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and chromatography (Isco Combi-Flash, 110g cartridge, 6:1 hexane-ethyl acetate) provided 207 (277 mg, 1.14 gtheoretical, 24%) as a colorless oil: R_(f) 0.32 (SiO₂, 6:1 hexane-ethylacetate); LRMS m/z 410 (M⁺+1, C₂₁H₂₂F₃NO₄ requires 410).

Example 175 Synthesis of 3-(4-Trifluoromethoxy-phenoxymethyl)-piperidine

A solution of 207 (0.677 mmol, 277 mg) and PdC 30% (50 mg) in CH₃OH (5mL) at 25° C. were added to a Paar hydrogenator low pressure reactionvessel. The mixture was reacted at 65 psi with vigorous shaking untilhydrogen uptake subsided (2 h). The catalyst was filtered through a padof celite. The filtrate was concentrated in vacuo which provided 208(125 mg, 186 mg theoretical, 67%) as colorless oil: LRMS m/z 276 (M⁺,C₁₃H₁₆F₃NO₂ requires 276).

Example 176 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl-piperidin-1-yl]-ethanone

A solution of 208 (0.301 mmol, 83 mg),2-bromo-1-[1-(4-chloro-phenyl)-cyclobutyl]-ethanone (1.4 equiv, 0.417mmol, 120 mg) and KF (50% wt on celite) (7.0 equiv, 2.11 mol. 245 mg) inCH₃CN (2 mL) was stirred for 12 h at 25° C. The reaction mixture wasfiltered, and the solvents were removed in vacuo. Chromatography (IscoCombi-Flash, 10 g cartridge, 2:1 hexane-ethyl acetate) provided 209 (98mg, 145 mg theoretical, 68%) as a colorless oil: R_(f) 0.46 (SiO₂, 2:1hexane-ethyl acetate); LRMS m/z 483 (M⁺+1, C₂₅H₂₇ClF₃NO₃ requires 483).

Example 177 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethoxy-phenoxymethyl-piperidin-1-yl]-ethanol

A solution of 209 (0.203 mmol, 98 mg) in CH₃OH (1 mL) was treated withNaBH₄ (1.5 equiv, 0.305 mmol, 12 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (10 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 10 g cartridge, 2:1 hexane-ethyl acetate) provided210 (93 mg, 98 mg theoretical, 95%) as a colorless oil: R_(f) 0.42(SiO₂, 2:1 hexane-ethyl acetate); LRMS m/z 485 (M⁺+1, C₂₅H₂₉ClF₃NO₃requires 485).

Example 178 Synthesis of2-[3-(4-Trifluoromethoxy-phenoxymethyl-piperidin-1-yl]-1-[1-(4-trifluoromethoxy-phenyl)-cyclobutyl]-ethanone

A solution of 208 (0.236 mmol, 65 mg), 204 (1.4 equiv, 0.323 mmol, 109mg) and KF (50% wt on celite) (7.0 equiv, 1.65 mol, 192 mg) in CH₃CN (2mL) was stirred for 12 h at 25° C. The reaction mixture was filtered,and the solvents were removed in vacuo. Chromatography (IscoCombi-Flash, 10 g cartridge, 2:1 hexane-ethyl acetate) provided 211 (101mg, 125 mg theoretical, 81%) as a colorless oil: R_(f) 0.52 (SiO₂, 2:1hexane-ethyl acetate); LRMS m/z 532 (M⁺+1, C₂₆H₂₇F₆NO₄ requires 532).

Example 179 Synthesis of2-[3-(4-Trifluoromethoxy-phenoxymethyl)-piperidin-1-yl]-1-[1-(4-trifluoromethoxy-phenyl)-cyclobutyl]-ethanol

A solution of 211 (0.190 mmol, 101 mg) in CH₃OH (1 mL) was treated withNaBH₄ (1.5 equiv, 0.285 mmol, 11 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (10 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 10 g cartridge, 2:1 hexane-ethyl acetate) provided212 (101 mg, 101 mg theoretical, 99%) as a colorless oil: R_(f) 0.49(SiO₂, 2:1 hexane-ethyl acetate); LRMS m/z 535 (M⁺+1, C₂₆H₂₉F₆NO₄requires 535).

Example 180 Synthesis of[1-(4-Chloro-phenyl-2-(3(R)-phenethyl-piperidin-1-yl)-ethyl]-methyl-amine

A solution of piperidine-phenyl ketone (0.493 mmol, 168 mg), CH₃NH₂ (2.0M in THF) (4.0 equiv, 1.97 mmol, 1 mL), NaCNBH₃ (4.0 equiv, 1.97 mmol,123 mg) in 5% AcOH in CH₃OH (2 mL) was stirred at 40° C. for 12 h. Thereaction mixture was quenched with 10% NaOH (10 mL) and extracted withEtOAc (2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and the residue waspurified by chromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm, 9:1CH₂C₁—H₃OH) which provided 213 (28 mg, 175 mg theoretical, 16%) as acolorless oil: R_(f) 0.35 (SiO₂, 9:1 CH₂ClCH₃OH); LRMS m/z 358 (M⁺+1,C₂₂H₂₉ClN₂ requires 358).

Example 181 Synthesis ofN-1-Carbobenzyloxy[3-S-(2′-anilino)carboxy]piperidine

A solution of S-Cbz-nipecotic acid (3.80 mmol, 1.00 g) and aniline (1.1equiv, 4.18 mmol, 381 μL) in CH₂Cl₂ (10 mL) at 0° C. was treated withDCC (1.5 equiv, 5.70 mmol, 1.18 g) under Ar. The reaction mixture wasallowed to warm to 25° C. and stirred for 12 h. The reaction mixture wasthen filtered to remove the urea and the solvents were removed in vacuo.Chromatography (Isco Combi-Flash, 110 g cartridge, 3:1 hexane-ethylacetate) provided the desired product (1.29 g, 1.29 g theoretical, 99%)as a white foam: R_(f) 0.45 (SiO₂, 1:1 hexane-ethyl acetate): LRMS m/z338 (M⁺+1, C₂₀H₂₂N₂O₃ requires 338).

Example 182 Synthesis of Piperidine-3-S-carboxilic Acid Phenylamide

A solution of 3-(S)-Phenylcarbamoyl-piperidine-1-carboxylic acid benzylester (1.80 mol, 608 mg) and Pd—C 30% (100 mg) in CH₃OH (10 mL) at 25°C. were added to a Paar hydrogenator low pressure reaction vessel. Themixture was reacted at 55 psi with vigorous shaking until hydrogenuptake subsided (2 h). The catalyst was filtered through a pad ofcelite. The filtrate was concentrated in vacuo which providedpiperidine-3-carboxylic acid phenylamide (367 mg, 367 mg theoretical,99%) as a white foam: LRMS m/z 205 (M⁺+1, C₁₂H₁₆N₂O requires 205).

Example 183 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutanecarbonyl]-piperidine-3-S-carboxylicAcid Phenylamide

A solution of the piperidine-3-carboxylic acid phenylamide (1.80 mmol,367 mg), 1-(4-Chlorophenyl)-1-cyclobutane carboxylic acid (1.2 equiv,2.16 mmol, 455 mg) and iPr₂NEt (3.0 equiv, 5.40 mmol, 0.941 mL) inCH₂Cl₂ (5 mL) was treated with BroP (1.5 equiv, 2.70 mmol, 2.07 g) underAr at 0° C. After warming to 25° C. and stirring for 12 h, the reactionmixture was quenched with 10% aqueous HCl and extracted with 3×EtOAc (25mL). The organic layer was then washed with NaHCO_(3(sat)) and driedwith NaCl_((sat)) and MgSO_(4(s)). Chromatography (Isco Combi-Flash, 35g cartridge, 3:2 hexane-ethyl acetate) provided 214 (632 mg, 714 mgtheoretical, 89%) as a white foam: R_(f) 0.17 (SiO₂, 2:1 hexane-ethylacetate); LRMS m/z 397 (M⁺+1, C₂₃H₂₅ClN₂O₂ requires 397).

Example 184 Synthesis of{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-S-ylmethyl}-phenyl-amine

A solution of 214 (0.504 mmol, 200 mg) in THF (2 mL) at 0° C. wastreated with LiAlH₄ (3.0 equiv, 1.51 mmol, 57 mg) under Ar. The reactionmixture stirred for 12 h and returned to 25° C. The reaction mixture wasthen cooled to 0° C., quenched with 10% aqueous NaOH and extracted with3×EtOAc (25 mL). The organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The reaction mixture was purified by chromatography (IscoCombi-Flash, 10 g cartridge, 8:2 hexane-ethyl acetate) which provided215 (88 mg, 186 mg theoretical, 47%) as a colorless oil: R_(f) 0.61(SiO₂, 2:1 hexane-ethyl acetate); LRMS m/z 369 (M⁺+1, C₂₃H₂₉ClN₂requires 369).

Example 185 Synthesis of{1-[1-(4-Chloro-phenyl)-cyclobutylymethyl]-piperidin-3-S-ylmethyl}-methyl-phenyl-amine

A solution of the 215 (0.238 mmol, 88 mg) in THF (1 mL) at −78° C. wastreated with 1.6 M nBuLi (1.5 equiv, 0.358 mmol, 224 μL) under Ar. Thereaction mixture was warmed to 0° C. for 30 min and then cooled again to−78° C. CH₃I (1.5 equiv, 0.358 mmol, 22 μL) was then added and thereaction mixture stirred at 0° C. for 5 min. The reaction was quenchedwith NaHCO_(3(sat)) and extracted with EtOAc. The combined organics weredried with NaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed invacuo and chromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm, 6:1hexanes-ethyl acetate) provided 216 (30 mg, 91 mg theoretical, 33%) as ayellow oil: R_(f) 0.38 (SiO₂, 3:1 hexanes-EtOAc); LRMS m/z 384 (M⁺+1,C₂₄H₃₁ClN₂ requires 384).

Example 186 Synthesis of1-(4-Chloro-phenyl)-2-methyl-3-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-propan-1-one

A solution of 208 (0.579 mmol, 150 mg),2-bromo-1-(4-chloro-phenyl)-2-methyl-propan-1-one (1.2 equiv, 0.695mmol, 182 mg) and KF (50% wt on celite) (7.0 equiv, 4.05 mol, 471 mg) inCH₃CN (2 mL) was stirred for 12 h at 25° C. The reaction mixture wasfiltered, and the solvents were removed in vacuo. Chromatography (IscoCombi-Flash, 10 g cartridge, 3:1 hexane-ethyl acetate) provided 217 (35mg, 255 mg theoretical, 14%) as a colorless oil: R_(f) 0.31 (SiO₂, 3:1hexane-ethyl acetate); LRMS m/z 441 (++1, C₂₃H₂₅ClF₃NO₂ requires 441).

Example 187 Synthesis of1-(4-Chloro-phenyl)-2-methyl-3-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-propan-1-ol

A solution of 217 (0.068 mmol, 30 mg) in CH₃OH (1 mL) was treated withNaBH₄ (4.0 equiv, 0.264 mmol, 10 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (10 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(PTLC, SiO₂, 20 cm×20 cm, 1 mm, 1:1 hexanes-ethyl acetate) provided 218(93 mg, 98 mg theoretical, 95%) as a colorless oil: R_(f) 0.38 (SiO₂,1:1 hexane-ethyl acetate); LRMS m/z 443 (M⁺+1, C₂₃H₂₇ClF₃NO₂ requires443).

Example 188 Synthesis of2-(5-Methoxy-1H-indol-3-yl)-1-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone

A solution of 208 (1.93 mmol, 500 mg) and(5-methoxy-1H-indol-3-yl)-acetic acid (1.2 equiv, 2.32 mmol, 476 mg) inCH₂Cl₂ (5 mL) was treated with BrOP (1.5 equiv, 2.90 mmol, 1.13 g) andiPr₂NEt (3.0 equiv, 5.79 mmol, 1.00 mL) at 0° C. The reaction mixturestirred for 12 h while warming to rt. The reaction mixture was quenchedwith 10% HCl (10 mL) and then extracted with EtOAc (2×15 mL). Thecombined organics were washed with NaHCO_(3(sat)) and dried withNaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed in vacuo andchromatography (Isco Combi-Flash, 110 g cartridge, 3:1 hexane-ethylacetate) provided 219 (617 mg, 862 mg theoretical, 72%) as a colorlessoil: R_(f) 0.33 (SiO₂, 3:1 hexane-ethyl acetate); LRMS m/z 447 (M⁺+1,C₂₄H₂₅F₃N₂O₃ requires 447).

Example 189 Synthesis of5-Methoxy-3-{2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethyl}-1H-indole

A solution of 219 (0.343 mmol, 153 mg) in THF (2 mL) at 0° C. wastreated with LiAlH₄ (3.0 equiv, 1.028 mmol, 39 mg) under Ar. Thereaction mixture stirred for 12 h and returned to 25° C. The reactionmixture was then cooled to 0° C., quenched with 10% aqueous NaOH andextracted with 3×EtOAc (25 mL). The organics were dried withNaCl_((sat)) and Na₂SO_(4(s)). The reaction mixture was purified bychromatography (Isco Combi-Flash, 10 g cartridge, 9:1 CH₂Cl₂CH₃OH) whichprovided 220 (128 mg, 148 mg theoretical, 86%) as a colorless oil: R_(f)0.16 (SiO₂, 9:1 CH₂Cl₂—CH₃OH); LRMS m/z 433 (M⁺+1, C₂₄H₂₇F₃N₂O₂)requires 433).

Example 190 Synthesis of1-(5-Chloro-1H-indol-3-yl-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanone

A solution of 208 (0.197 mmol, 51 mg),2-chloro-1-(5-chloro-1H-indol-3-yl)-ethanone (1.0 equiv, 0.197 mmol, 45mg) and KF (50% wt on celite) (7.0 equiv, 1.38 mol, 160 mg) in CH₃CN (2mL) was stirred for 12 h at 25° C. The reaction mixture was filtered,and the solvents were removed in vacuo. Chromatography (IscoCombi-Flash, 10 g cartridge, 3:1 hexane-ethyl acetate) provided 221 (53mg, 89 mg theoretical, 60%) as a colorless oil: R_(f) 0.34 (SiO₂, 3:1hexane-ethyl acetate); LRMS m/z 451 (M⁺+1, C₂₃H₂₂F₃N₂O₂ requires 451).

Example 191 Synthesis of1-(5-Chloro-1H-indol-3-yl)-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A solution of 221 (0.118 mmol, 53 mg) in CH₃OH (1 mL) was treated withNaBH (3.0 equiv, 0.354 mmol, 13 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (10 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_(4(sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 10 g cartridge, 2:1 hexane-ethyl acetate) provided222 (23 mg, 53 mg theoretical, 43%) as a colorless oil: R_(f) 0.33(SiO₂, 2:1 hexane-ethyl acetate); LRMS m/z 454 (M⁺+1, C₂₃H₂₄F₃N₂O₂requires 454).

Example 192 Synthesis of1-[1-(2-Trifluoromethoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxy-R-methyl)-piperidin-1-yl]-ethanone

A solution of 74 (0.683 mmol, 177 mg),2-bromo-1-[1-(2-trifluoromethoxy-phenyl0-cyclobutyl]-ethanone (1.0equiv, 0.683 mmol, 200 mg) and KF (50% wt on celite) (7.0 equiv, 4.78mol, 560 mg) in CH₃CN (4 mL) was stirred for 12 h at 25° C. The reactionmixture was filtered, and the solvents were removed in vacuo.Chromatography (Isco Combi-Flash, 25 g cartridge, 3:1 hexane-ethylacetate) provided 223 (193 mg, 352 mg theoretical, 55%) as a colorlessoil: R_(f) 0.55 (SiO₂, 2:1 hexane-ethyl acetate); LRMS m/z 516 (M+1,C₂₆H₂₇F₆NO₃ requires 516).

Example 193 Synthesis of1-[1-(2-Trifluoromethoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol;Preparation of Diastereomers

A solution of 223 (0.374 mmol, 193 mg) in CH₃OH (1 mL) was treated withNaBH₄ (3.0 equiv, 1.12 mmol, 42 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (10 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 10 g cartridge, 2:1 hexane-ethyl acetate) provided224 and 225 (174 mg, 194 mg theoretical, 90%) as a colorless oil: R_(f)0.48 (SiO₂, 2:1 hexane-ethyl acetate); LRMS m/z 518 (M⁺+1, C₂₆H₂₉F₆NO₃requires 518). The two diastereomers were then separated on a ChiracelOD column: 224 and 225 were dissolved in hexane at a concentration of 85mg/mL; and the compounds were separated by using a 99.5% hexane and 0.5%isopropanol solvent system providing 224 (first peak) and 225 (secondpeak).

Example 194 Synthesis of1-[1-(2-Trifluoromethoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxy-3S-methyl)-piperidin-1-yl]-ethanone

A solution of 3(S)-(4-trifluoromethyl-phenoxymethyl)-piperidine (0.683mmol, 177 mg),2-bromo-1-[1-(2-trifluoromethoxy-phenyl0-cyclobutyl]-ethanone (1.0equiv, 0.683 mmol, 200 mg) and KF (50% wt on celite) (7.0 equiv, 4.78mol, 556 mg) in CH₃CN (4 mL) was stirred for 12 h at 25° C. The reactionmixture was filtered, and the solvents were removed in vacuo.Chromatography (Isco Combi-Flash, 25 g cartridge, 3:1 hexane-ethylacetate) provided 226 (160 mg, 352 mg theoretical, 45%) as a colorlessoil: R_(f) 0.55 (SiO₂, 2:1 hexane-ethyl acetate); LRMS m/z 516 (M⁺+1,C₂₆H₂₇F₆NO₃ requires 516).

Example 195 Synthesis of1-[1-(2-Trifluoromethoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol

A solution of 226 (0.374 mmol, 193 mg) in CH₃OH (1 mL) was treated withNaBH₄ (3.0 equiv, 1.12 mmol, 42 mg) at 0° C. The reaction mixture wasallowed to warm to rt and stirred for 12 h. The reaction mixture wasquenched with pH 7 phosphate buffer (10 mL) and extracted with EtOAc(2×10 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo and chromatography(Isco Combi-Flash, 10 g cartridge, 2:1 hexane-ethyl acetate) provided227 and 228 (140 mg, 160 mg theoretical, 88%) as a colorless oil: R_(f)0.48 (SiO₂, 2:1 hexane-ethyl acetate); LRMS m/z 518 (M⁺+1, C₂₆H₂₉F₆NO₃requires 518). The two diastereomers were separated on a Chiracel ODcolumn: 227 and 228 were dissolved in hexane at a concentration of 70mg/mL; and the compounds were separated by using a 99.5% hexane and 0.5%isopropanol solvent system providing 227 (first peak) and 228 (secondpeak).

Example 196 Synthesis of(3-Azidomethyl-piperidin-1-yl)-[1-(4-chloro-phenyl)-cyclobutyl]-methanone

A solution of methanesulfonic acid1-[1-(4-chloro-phenyl)-cyclobutanecarbonyl]-piperidin-3-ylmethyl ester(2.46 mmol, 950 mg) and NaN₃ (10 equiv, 24.6 mmol, 1.60 g) in DMF (10mL) was heated at 70° C. for 48 h. The reaction mixture was quenchedwith H₂O (50 mL) and then extracted with EtOAc (2×50 mL). The combinedorganics were dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (Isco Combi-Flash, 110 gcartridge, 2:1 hexane-ethyl acetate) provided 229 (725 mg, 819 mgtheoretical, 89%) as a pale yellow oil: R_(f) 0.68 (Sir, 1:1hexane-ethyl acetate); LRMS m/z 333 (M⁺+1, C₁₇H₂₃ClN₄O requires 333).

Example 197 Synthesis ofC-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-yl}-methylamine

A solution of 229 (0.225 mmol, 75 mg) in THF (1 mL) at 0° C. was treatedwith LiAlH₄ (3.0 equiv, 0.676 mmol, 26 mg) under Ar. The reactionmixture stirred at 60° C. for 12 h. The reaction mixture was then cooledto 0° C., quenched with 10% aqueous NaOH and extracted with 2×EtOAc (15mL). The organics were dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed to provide 230 (57 mg, 66 mg theoretical, 86%) asa colorless oil: LRMS m/z 293 (M⁺+1, C₁₇H₂₅ClN₂ requires 293).

Example 198 Synthesis of2-[3-(benzo[1,3]dioxol-5-yloxymethyl)-piperidin-1-yl]-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanol

2-[3-(Benzo[1,3]dioxol-5-yloxymethyl)-piperidin-1-yl]-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanone(ketone not shown) was prepared according to the procedures outlined inExample 138, using 231 (150 mg, 0.638 mmol), KF-Celite (50% weight onCelite; 520 mg, 0.638 mmol), and2-chloro-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanone (152 mg, 0.638mmol) in acetonitrile (4 mL); yield: 71 mg; MS⁺ (437).

2-[3-(Benzo[1,3]dioxol-5-yloxymethyl)-piperidin-1-yl]-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanol(232) was prepared according to the procedures outlined in Example 138,using sodium borohydride (17 mg, 0.453 mmol), and2-[3-(benzo[1,3]dioxol-5-yloxymethyl)-piperidin-1-yl]-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanone(100 mg, 0.227 mmol) in MeOH (1.6 mL); MS⁺(440).

Example 199 Synthesis of the Individual Stereoisomers of1-[1-(2-Methoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanolPreparation of Chiral Epoxides

Following the same procedures used for the asymmetric reduction of2-Bromo-1-[1-(4-chloro-phenyl)-cyclobutyl]-ethanone (See Example 127),500 mg samples of 2-Chloro-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanonewere reduced to the enantiomerically enriched alcohols withborane-methyl sulfide in the presence of S-2-methyl CBS-oxazaborolidineor R-2-methyl CBS-oxazaborolidine. Each isomer of2-Chloro-1-[1-(2-methoxy-phenyl)-cyclobutyl]-ethanol was obtained inapproximately 70% yield and ca. 90% e.e. based on chiral HPLC analysis.240 mg of each chloroalcohol was converted to the corresponding epoxideusing 1.1 equivalents of cesium carbonate in 5 mL of THF at 50° C. for 3hours. Dilution with 20 mL of water, addition of 30 mL ether, andextractive workup gave in each instance, after concentration of theorganic layers in vacuo and chromatography on silica gel, each of thedesired epoxides (176 mg and 184 mg, 86% and 90% yield).

Preparation of Individual Stereoisomers

50 mg of R-2-[1-(2-Methoxy-phenyl)-cyclobutyl]-oxirane and 63 mg ofR-3-(4 Trifluoromethyl-phenoxymethyl)-piperidine in 4 mL of methanolwere heated in a sealed tube at 100° C. for 16 hours, cooled to roomtemperature, transferred to a round-bottomed flask, and concentrated invacuo. The crude residue was purified by preparative HPLC using aChiralpak AD™ column from Chiral Technologies, Inc., eluting with a 98:2mixture of hexane and isopropyl alcohol containing ca. 0.1%diethylamine. 43 mg of the R,R′ isomer of1-[1-(2-Methoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol(236) was obtained (38%).

In the same fashion, 50 mg ofR-2-[1-(2-Methoxy-phenyl)-cyclobutyl]-oxirane and 63 mg ofS-3-(4-Trifluoromethyl-phenoxymethyl)-piperidine gave the S,R′ isomer of1-[1-(2-Methoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol(234) (35%), 50 mg of S-2-[1-(2-Methoxy-phenyl)-cyclobutyl]-oxirane and63 mg of R-3-(4-Trifluoromethyl-phenoxymethyl)-piperidine gave the R,S′isomer of1-[1-(2-Methoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol(235) (40%), and 50 mg of S-2-[1-(2-Methoxy-phenyl)-cyclobutyl]-oxiraneand 63 mg of S-3-(4-Trifluoromethyl-phenoxymethyl)-piperidine gave theS,S′ isomer of1-[1-(2-Methoxy-phenyl)-cyclobutyl]-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-ethanol(237) (41%). ¹H and ¹³C (DEPT) NMR data for each isomer was consistentwith the assigned structure. Data for each diastereomer: MS 464 (M⁺+1).

Example 200 Synthesis of 2-(4-Chloro-phenyl)-1-(3(S)-phenethyl-piperidin-1-yl)-propan-2-ol

A solution of (S)-piperidine-ketone (0.292 mmol, 100 mg) in THF (1 mL)was added to a solution of CH₃MgCl (1.0 M in THF) (1.5 equiv, 0.44 mmol,0.44 mL) in THF (1 mL) at 0° C. The reaction mixture stirred for 1 h.The reaction mixture was quenched with 10% NaOH (10 mL) and extractedwith EtOAc (2×10 mL). The combined organics were dried with NaCl_((sat))and Na₂SO_(4(s)). The solvents were removed in vacuo and the residue waspurified by chromatography (PTLC, SiO₂, 20 cm×20 cm, 1 mm, 3:1hexane-acetone) which provided desired product (40 mg, 38%) as acolorless oil: R_(f) 0.44 (SiO₂, 3:1 hexane-acetone); LRMS m/z 359(M⁺+1, C₂₂H₂₈ClNO requires 359).

Example 201 Synthesis of R-3-Hydroxymethyl-pyrrolidine-1-carboxylic AcidBenzyl Ester

A solution of (−)-2-pyrrolidinemethanol (20 mmol, 2.0 g), CbzCl (2.0equiv, 40 mmol, 6 mL) and K₂CO₃ (1.5 equiv, 30 mmol, 4.1 g) in THF/H₂O(1:1) (66 mL) at 0° C. was allowed to warm to 25° C. and stirred for 12h. Ethyl acetate (100 mL) was added and the layers were separated. Theorganic layer was washed with 10% aqueous HCl and then washed withNaHCO_(3(sat)) and dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and chromatography (Isco Combi-Flash, 110g cartridge, 1:1 Hexane-EtOAc) provided 240 (3.74 g, 4.71 g theoretical,79%) as a colorless oil: LRMS m/z 236 (M⁺+1, C₁₃H₁₇NO₃, requires 236).

Example 202 Synthesis of S-3-Hydroxymethyl-pyrrolidine-1-carboxylic AcidBenzyl Ester

A solution of (−)-2-pyrrolidinemethanol (20 mmol, 2.0 g), CbzCl (2.0equiv, 40 mmol, 6 mL) and K₂CO₃ (1.5 equiv, 30 mmol, 4.1 g) in THF/H₂O(1:1) (66 mL) at 0° C. was allowed to warm to 25° C. and stirred for 12h. Ethyl acetate (100 mL) was added and the layers were separated. Theorganic layer was washed with 10% aqueous HCl and then washed withNaHCO_(3(sat)) and dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and chromatography (Isco Combi-Flash, 110g cartridge, 1:1 Hexane-EtOAc) provided 241 (4.38 g, 4.71 g theoretical,93%) as a colorless oil: LRMS m/z 236 (M⁺+1, C₁₃H₁₇NO₃, requires 236).

Example 203 Synthesis ofR-3-Methanesulfonyloxymethyl-pyrrolidine-1-carboxylic Acid Benzyl Ester

A solution of 240 (4.2 mmol, 1.0 g), MsCl (1.5 equiv, 6.4 mmol, 0.5 mL)and iPr₂NEt (1.5 equiv, 6.4 mmol, 1.1 mL) in CH₂Cl₂ (14 mL) at 0° C. wasallowed to warm to 25° C. and stirred for 12 h. The reaction mixture wasquenched with 10% HCl (20 mL) and then extracted with EtOAc (2×50 mL).The combined organics were dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and chromatography (Isco Combi-Flash, 110g cartridge, 1:1 Hexane-EtOAc) provided 242 as a colorless oil: LRMS m/z314 (M⁺+1, C₁₄H₁₉NO₅S, requires 314).

Example 204 Synthesis ofS-3-Methanesulfonyloxymethyl-pyrrolidine-1-carboxylic Acid Benzyl Ester

A solution of 241 (4.2 mmol, 1.0 g), MsCl (1.5 equiv, 6.4 mmol, 0.5 mL)and iPr₂NEt (1.5 equiv, 6.4 mmol, 1.1 mL) in CH₂Cl₂ (14 mL) at 0° C. wasallowed to warm to 25° C. and stirred for 12 h. The reaction mixture wasquenched with 10% HCl (20 mL) and then extracted with EtOAc (2×50 mL).The combined organics were dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and chromatography (Isco Combi-Flash, 110g cartridge, 1:1 Hexane-EtOAc) provided 243 as a colorless oil: LRMS m/z314 (M⁺+1, C₁₄H₁₉NO₅S, requires 314).

Example 205 Synthesis ofR-3-(4-Trifluoromethyl-phenoxymethyl)-pyrrolidine-1-carboxylic AcidBenzyl Ester

A solution of 242 (3.8 mmol, 1.2 g), α,α,α-trifluoro-p-cresol (1.5equiv, 5.7 mmol, 0.9 g) and Cs₂CO₃ (2.0 equiv, 7.6 mmol, 2.5 g) in CH₃CN(13 mL) was heated to 90° C. and stirred for 12 h. Ethyl acetate (100mL) and H₂O (100 mL) were added and the layers were separated. Theorganic layer was dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (Isco Combi-Flash, 110 gcartridge, 9:1 Hexane-EtOAc) provided 244 (0.762 g, 1.44 g theoretical,53%) as a colorless oil: LRMS m/z 380 (++1, C₂₀H₂₀F₃NO₃, requires 380).

Example 206 Synthesis ofS-3-(4-Trifluoromethyl-phenoxymethyl)-pyrrolidine-1-carboxylic AcidBenzyl Ester

A solution of 243 (3.8 mmol, 1.2 g), α,α,α-trifluoro-p-cresol (1.5equiv, 5.7 mmol, 0.9 g) and Cs₂CO₃ (2.0 equiv, 7.6 mmol, 2.5 g) in CH₃CN(13 mL) was heated to 90° C. and stirred for 12 h. Ethyl acetate (100mL) and H₂O (100 mL) were added and the layers were separated. Theorganic layer was dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (Isco Combi-Flash, 110 gcartridge, 9:1 Hexane-EtOAc) provided 245 (0.421 g, 1.44 g theoretical,29%) as a colorless oil: LRMS m/z 380 (M⁺1, C₂₀H₂₀F₃NO₃, requires 380).

Example 207 Synthesis ofR-3-(4-Trifluoromethyl-phenoxymethyl)-pyrrolidine

A solution of 244 (1.3 mmol, 486 mg) in CH₃OH (13 mL) was treated 30%Pd—C (81 mg) and H₂ (Hydrogen balloon). The reaction was stirred for 5h. The reaction mixture was filtered through celite, and the solventswere removed in vacuo to provide 246 (319 mg, 319 mg theoretical,quantitative) as a colorless oil: LRMS m/z 246 (M⁺+1, C₁₂H₁₄F₃NO,requires 246).

Example 208 Synthesis ofS-3-(4-Trifluoromethyl-phenoxymethyl)-pyrrolidine

A solution of 245 (1.1 mmol, 421 mg) in CH₃OH (11 mL) was treated 30%Pd—C (70 mg) and H₂ (Hydrogen balloon). The reaction was stirred for 5h. The reaction mixture was filtered through celite, and the solventswere removed in vacuo to provide 247 (270 mg, 270 mg theoretical,quantitative) as a colorless oil: LRMS m/z 246 (M⁺1, C₁₂H₁₄F₃NO,requires 246).

Example 209 Synthesis ofR-[3-(4-Trifluoromethyl-phenoxymethyl)-pyrrolidin-1-yl]-[1-(4-trifluoromethyl-phenyl)-cyclobutyl]-methanone

A solution of the 246 (1.3 mmol, 310 mg),1-(4-Chlorophenyl)-1-cyclobutane carboxylic acid (1.5 equiv, 1.9 mmol,400 mg) and iPr₂NEt (3.0 equiv, 4.0 mmol, 0.7 mL) in CH₂Cl₂ (5 mL) wastreated with PyBroP (1.5 equiv, 1.9 mmol, 890 mg) under Ar at 0° C.After warming to 25° C. and stirring for 12 h, the reaction mixture wasquenched with 10% aqueous HCl and extracted with 3×EtOAc (25 mL). Theorganic layer was then washed with NaHCO_(3(sat)) and dried withNaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed in vacuo andchromatography (Isco Combi-Flash, 110 g cartridge, 3:1 Hexane-EtOAc)provided 248 (406 mg, 569 mg theoretical, 71%) as a colorless oil: LRMSm/z 439 (M⁺+1, C₂₃H₂₃ClF₃NO₂, requires 439).

Example 210 Synthesis ofS-[3-(4-Trifluoromethyl-phenoxymethyl)-pyrrolidin-1-yl]-[1-(4-trifluoromethyl-phenyl)-cyclobutyl]-methanone

A solution of the 247 (1.1 mmol, 280 mg),1-(4-Chlorophenyl)-1-cyclobutane carboxylic acid (1.5 equiv, 1.7 mmol,358 mg) and iPr₂NEt (3.0 equiv, 3.4 mmol, 0.6 mL) in CH₂Cl₂ (4 mL) wastreated with PyBroP (1.5 equiv, 1.7 mmol, 792 mg) under Ar at 0° C.After warming to 25° C. and stirring for 12 h, the reaction mixture wasquenched with 10% aqueous HCl and extracted with 3×EtOAc (25 mL). Theorganic layer was then washed with NaHCO_(3(sat)) and dried withNaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed in vacuo andchromatography (Isco Combi-Flash, 110 g cartridge, 3:1 Hexane-EtOAc)provided 249 (230 mg, 482 mg theoretical, 48%) as a colorless oil: LRMSm/z 439 (M⁺+1, C₂₃H₂₃ClF₃NO₂, requires 439).

Example 211 Synthesis ofR-3-(4-Trifluoromethyl-phenoxymethyl)-1-[1-(4-trifluoromethyl-phenyl)-cyclobutylmethyl]-pyrrolidine

A solution of 248 (0.23 mmol, 100 mg) in THF (1 mL) at 25° C. wastreated with LiAlH₄ (3.0 equiv, 0.69 mmol, 26 mg) under Ar. The reactionmixture stirred for 12 h at 60° C. The reaction mixture was then cooledto 0° C., quenched with 10% aqueous NaOH and extracted with 3×EtOAc (25mL). The organics were dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and chromatography (Isco Combi-Flash, 25g cartridge, 3:1 Hexane-EtOAc) provided 250 (65 mg, 97 mg theoretical,67%) as a colorless oil: LRMS m/z 425 (M⁺+1, C₂₃H₂₅ClF₃NO, requires425).

Example 212 Synthesis ofS-3-(4-Trifluoromethyl-phenoxymethyl)-1-[1-(4-trifluoromethyl-phenyl)-cyclobutylmethyl]-pyrrolidine

A solution of 249 (0.23 mmol, 100 mg) in THF (1 mL) at 25° C. wastreated with LiAlH₄ (3.0 equiv, 0.69 mmol, 26 mg) under Ar. The reactionmixture stirred for 12 h at 60° C. The reaction mixture was then cooledto 0° C., quenched with 10% aqueous NaOH and extracted with 3×EtOAc (25mL). The organics were dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and chromatography (Isco Combi-Flash, 25g cartridge, 3:1 Hexane-EtOAc) provided 251 (73 mg, 97 mg theoretical,75%) as a colorless oil: LRMS m/z 425 (M⁺+1, C₂₃H₂₅ClF₃NO, requires425).

Example 213 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-(3-S-phenethyl-piperidin-1-yl)-S-ethanol

A solution of the amine (1.2 equiv, 1.58 mmol, 300 mg) and the epoxide(1.32 mmol, 275 mg) in CH₃CN (2 mL) was stirred for 12 h at 95° C. Thereaction mixture was quenched with H₂O and extracted with 3×EtOAc (25mL). The organics were dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and chromatography (Isco Combi-Flash, 10g cartridge, 1:1 Hexane-EtOAc) provided 252 (439 mg, 525 mg theoretical,84%) as a colorless oil: R_(f) 0.42 (SiO₂, 1:1 hexamethyl acetate); LRMSm/z 399 (M⁺+1, C₂₅H₃₂ClNO, requires 399).

Example 214 Synthesis of1-[1-(4-Chloro-phenyl)-cyclobutyl]-2-(3-R-phenethyl-piperidin-1-yl)-R-ethanol

A solution of (R)-3-phenethyl piperidine (1.0 equiv, 2.14 mmol, 405 mg),alcohol (2.14 mmol, 619 mg) and K₂CO₃ (1.5 equiv, 3.21 mmol, 444 mg), inCH₃CN (2 mL) was stirred for 12 h at 95° C. in a sealed pressure tube.The reaction mixture was quenched with H₂O and extracted with 3×EtOAc(25 mL). The organics were dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and chromatography (Isco Combi-Flash, 25g cartridge, Hexane-EtOAc (45%)) provided 253 (852 mg, 760 mgtheoretical, 89%) as a colorless oil: R_(f) 0.38 (SiO₂, hexane-ethylacetate (45%)); LRMS m/z 399 (M⁺+1, C₂₅H₃₂ClNO, requires 399).

Example 215 Synthesis ofR-(3-Azidomethyl-piperidin-1-yl)-[1-(4-chloro-phenyl)-cyclobutyl]-methanone

A solution of the mesylate (2.29 mmol, 884 mg) and NaN₃ (10 equiv, 22.9mmol, 1.49 g) in DMF (10 mL) was heated at 70° C. for 48 h. The reactionmixture was quenched with H₂O (50 mL) and then extracted with EtOAc(2×50 mL). The combined organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed in vacuo which provided 254 (762mg, 650 mg theoretical, 85%) as a pale yellow oil: R_(f) 0.68 (SiO₂, 1:1hexane-ethyl acetate); LRMS m/z 333 (M⁺+1, C₁₇H₂₃ClN₄O requires 333).

Example 216 Synthesis ofC-{1-[1-(4-Chloro-phenyl)-cyclobutylmethyl]-piperidin-3-R-yl}-methylamine

A solution of 254 (0.451 mmol, 150 mg) in THF (2 mL) at 0° C. wastreated with LiAlH₄ (3.0 equiv, 1.35 mmol, 51 mg) under Ar. The reactionmixture stirred for 12 h and returned to 60° C. The reaction mixture wasthen cooled to 0° C., quenched with 10% aqueous NaOH and extracted with2×EtOAc (25 mL). The organics were dried with NaCl_((sat)) andNa₂SO_(4(s)). The solvents were removed to provide 255 as a colorlessoil: LRMS m/z 293 (M⁺+1, C₁₇H₂₅ClN₂ requires 293).

Example 217 Synthesis of 3-Iodomethyl-Piperidine-1-Carboxylic AcidBenzyl Ester

A solution of triphenyl phosphine (1.5 equiv, 30 mmol, 7.87 g) andimidazole (1.5 equiv, 30 mmol, 2.05 g) in CH₂Cl₂ (50 mL) at 0° C. wastreated with 12 (1.5 equiv, 30 mmol, 7.61 g). After 5 min, the alcohol(20 mmol, 5.00 g) in CH₂Cl₂ (10 mL) was added at 0° C. The reactionstirred for 1 h at 25. The reaction mixture was quenched with 10%aqueous HCl and extracted with 3×EtOAc (75 mL). The organics were washedwith H₂O then dried with NaCl_((sat)) and Na₂SO_(4(s)). The solventswere removed in vacuo and chromatography (Isco Combi-Flash, 110 gcartridge, 7:1 Hexane-EtOAc) provided the desired compound (5.90 g, 7.18g theoretical, 82%) as a colorless oil: LRMS m/z 360 (M⁺+1, C₁₄H₁₈INO₂,requires 360).

Example 218 Synthesis of2-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidine-1-carbonyl]-morpholine-4-carboxylicAcid Tert-Butyl Ester

A solution of the amine (1.93 mmol, 500 mg), morpholine-2,4-dicarboxylicacid 4-tert-butyl ester (1.1 equiv, 2.12 mmol, 490 mg) and iPr₂NEt (3.0equiv, 5.79 mmol, 1.00 mL) in CH₂Cl₂ (10 mL) was treated with BroP (1.5equiv, 2.90 mmol, 1.13 g) under Ar at 0° C. After warming to 25° C. andstirring for 12 h, the reaction mixture was quenched with 10% aqueousHCl and extracted with 3×EtOAc (25 mL). The organic layer was thenwashed with NaHCO_(3(sat)) and dried with NaCl_((sat)) and Na₂SO_(4(s)).The solvents were removed in vacuo and chromatography (Isco Combi-Flash,110 g cartridge, 1:1 Hexane-EtOAc) provided the desired compound (632mg, 912 mg theoretical, 69%) as a colorless oil: LRMS m/z 474 (M⁺+1,C₂₃H₃₁F₃N₂O₅, requires 474).

Example 219 Synthesis ofmorpholin-2-yl-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-yl]-methanone

A solution of the amide (1.93 mmol, 500 mg) in CH₂Cl₂ (0.5 mL) wastreated with TFA (0.5 mL) under Ar at 0° C. After stirring for 1 h, thesolvents were removed in vacuo, and the residue was dissolved in EtOAc(25 mL). The organic layer was then washed with NaHCO_(3(sat)) and driedwith NaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed in vacuowhich provided 256 (74 mg, 74 mg theoretical, quantitative) as acolorless oil: LRMS m/z 373 (M⁺+1, C₁₈H₂₃F₃N₂O₃, requires 373).

Example 220 Synthesis of2-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidin-1-ylmethyl]-morpholine-4-carboxylicAcid Benzyl Ester

A solution of the amide (0.849 mmol, 316 mg) in THF (2 mL) at 0° C. wastreated with LiAlH₄ (3.0 equiv, 2.55 mmol, 97 mg) under Ar. The reactionmixture stirred for 12 h and returned to 60° C. The reaction mixture wasthen cooled to 0° C., quenched with 10% aqueous NaOH and extracted with2×EtOAc (25 mL). The organics were dried with NaCl_(4(sat)) andNa₂SO_(4(s)). After removing the solvents, the residue was dissolved inTHF/H₂O (3 mL) and treated with K₂CO₃ (3.0 equiv, 2.55 mmol 352 mg) andCbzCl (1.5 equiv, 1.27 mmol, 182 μL). The reaction mixture was quenchedwith H₂O and extracted with 3×EtOAc (10 mL). The organics were driedwith NaCl_((sat)) and Na₂SO_(4(s)). The solvents were removed in vacuoand chromatography (Isco Combi-Flash, 25 g cartridge, CH₂Cl₂-CH₃OH(10%)) provided the desired compound (166 mg, 418 mg theoretical, 40%)as a colorless oil: LRMS m/z 494 (M⁺+1, C₂(H₃₁F₃N₂O₄, requires 494).

Example 221 Synthesis of2-[3-(4-Trifluoromethyl-phenoxymethyl)-piperidin-1-ylmethyl]-morpholine

A solution of the amine (0.337 mmol, 166 mg) in CH₃OH (2 mL) was treated30% PdC (50 mg) and H₂ (Hydrogen balloon). The reaction was stirred for5 h. The reaction mixture was filtered through celite, and the solventswere removed in vacuo to provide 257 (114 mg, 121 mg theoretical, 94%)as a colorless oil: LRMS m/z 359 (M⁺+1, C₁₈H₂₅F₃N₂O₂, requires 359).

Example 222 Synthesis of4-(4-Chloro-phenyl-2-[3-(4-trifluoromethyl-phenoxymethyl)-piperidin-1-ylmethyl]-morpholine

A solution of the amine (0.318 mmol, 114 mg), 1-bromo-4-chloro-benzene(1.0 equiv, 0.318 mmol, 61 mg), BINAP (4.0 mol %, 0.0127 mmol, 8 mg) andNaOtBu (1.4 equiv, 0.445 mmol, 43 mg) in toluene (0.5 mL) was treatedwith Pd₂(DBA)₃ (2.0 mol %, 0.00636 mmol, 6 mg) under Ar at 0° C. Thereaction was heated to 70° C. for 12 h. The reaction mixture wasquenched with pH 7 buffer solution and extracted with 3×EtOAc (10 mL).The organic layer was then dried with NaCl_((sat)) and Na₂SO_(4(s)). Thesolvents were removed in vacuo and chromatography (Isco Combi-Flash, 10g cartridge, 1:1 Hexane-EtOAc) provided 258 (100 mg, 149 mg theoretical,67%) as a colorless oil: R_(f) 0.37 (SiO₂, 1:1 hexane-ethyl acetate);LRMS m/z 470 (M⁺+1, C₂₄H₂₈ClF₃N₂O₂, requires 470).

Example 223 Spontaneous Locomotor Activity in Rats

The effect of 124 and 126 on spontaneous locomotor activity in rats wasdetermined according to the procedures outlined by Silverman et al.(Motor Activity. In “Animal behavior in the laboratory”, Chapman andHall eds, London, p. 79-92, 1978) and Boissier et al. (Arch. Int.Pharmacodyn. 1965, 158, 212.)

Test items and test item vehicles were administered to maleSprague-Dawley rats (n=10) as a single i.p. dose. One, three, five,eight and twenty four hours following administration, rats were placedin a plastic box 30×30 cm in a room with low light intensity (maximum 50lux). Locomotor activity was determined during 20 minute periods usingvideo image analyzers. Images recorded with video cameras weredigitalized and displacements of the center of gravity of the digitalimage spot were tracked and analyzed. When the speed of displacement ofthe center of gravity of the spot was below 4.26 cm/sec, the movementwas considered as inactivity. When this speed was between 4.26 and 6.75cm/sec, the movement was considered as a small movement. When this speedwas above 6.75 cm/sec, the movement was considered as a large movement.The number of occurrences, distance and duration of fast and slowmovements, number of occurrences and duration of periods of inactivityand number of rears were measured.

Compounds 124 and 126, when dosed at 10 mg/kg, showed a significantincrease in locomotor activity compared to control animals at all timestested.

Methyl- 126 124 phenidate, 10 mg/kg 10 mg/kg 10 mg/kg (Dosed in (Dosedin (Dosed in Vehicle A Vehicle B Vehicle A) Vehicle B) Vehicle B) Large1 hour:  1 hour:  1 hour:  1 hour:  1 hour:  Movement 294 ± 47 236 ± 50 774 ± 110 655 ± 83  958 ± 118 Occurrences 3 hours: 3 hours: 3 hours: 3hours: 3 hours: 108 ± 25 139 ± 32 956 ± 93  751 ± 122  504 ± 114 5hours: 5 hours: 5 hours: 5 hours: 5 hours: 114 ± 56  68 ± 26  866 ± 117616 ± 82 196 ± 62 8 hours: 8 hours: 8 hours: 8 hours: 8 hours:  64 ± 26 89 ± 18 746 ± 95 455 ± 69  71 ± 20 24 hours:  24 hours:  24 hours:  24hours:  24 hours:  215 ± 38 124 ± 30 1012 ± 81  440 ± 55 335 ± 68 SmallMovement 1 hour:  1 hour:  1 hour:  1 hour:  1 hour:  Occurrences 815 ±86 661 ± 96 1357 ± 125 1258 ± 83  1595 ± 83  3 hours: 3 hours: 3 hours:3 hours: 3 hours: 488 ± 77 564 ± 77 1610 ± 73  1386 ± 96  1072 ± 122 5hours: 5 hours: 5 hours: 5 hours: 5 hours:  461 ± 108 329 ± 99 1575 ±90  1277 ± 100  612 ± 108 8 hours: 8 hours: 8 hours: 8 hours: 8 hours:309 ± 80 441 ± 59 1489 ± 111 1042 ± 109 358 ± 65 24 hours:  24 hours: 24 hours:  24 hours:  24 hours:  642 ± 82 486 ± 71 1544 ± 75  934 ± 87790 ± 81

INCORPORATION BY REFERENCE

All of the patents and publications cited herein are hereby incorporatedby reference.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

1. A compound represented by A:

wherein X is C(R₃)₂; Z is O; k is 1 or 2; n is 1; y is 1; R₁ is aryl; R₃is H; R₄ is cycloalkyl, aryl, or heteroaryl; R₅ and R₆ are eachindependently H, alkyl, or OH; R₅′ and R₆′ are each independently alkyl;R₅′ and R₆′ are connected through a covalent bond to form a ring; R₈ andR₉ are H; and the stereochemical configuration at any stereocenter of acompound represented by A is R, S, or a mixture of these configurations.2-125. (canceled)
 126. The compound of claim 1, wherein said compound isa single stereoisomer.
 127. A formulation, comprising a compound ofclaim 1, and a pharmaceutically acceptable excipient. 128-157.(canceled)
 158. A method of treating addiction, anxiety, depression,sexual dysfunction, hypertension, migraine, Alzheimer's disease,obesity, emesis, psychosis, analgesia, schizophrenia, Parkinson'sdisease, restless leg syndrome, sleeping disorders, attention deficithyperactivity disorder, irritable bowel syndrome, premature ejaculation,menstrual dysphoria syndrome, urinary incontinence, inflammatory pain,neuropathic pain, Lesche-Nyhane disease, Wilson's disease, or Tourette'ssyndrome, comprising the step of: administering to a patient atherapeutically effective amount of a compound of claim
 1. 159.(canceled)
 160. The method of claim 158, wherein the patient is a human.161. The method of claim 158, wherein the compound is administeredorally.
 162. The method of claim 158, wherein the compound isadministered intravenously. 163-164. (canceled)
 165. The method of claim158, wherein the compound is administered transdermally. 166-168.(canceled)
 169. The method of claim 158, wherein the compound isadministered intramuscularly.
 170. The method of claim 158, wherein thecompound is administered subcutaneously. 171-172. (canceled)
 173. Thecompound of claim 1, wherein R₁ is 4-trifluoromethylphenyl or3,4-methylenedioxyphenyl.
 174. The compound of claim 1, wherein R₄ is4-chlorophenyl.
 175. The compound of claim 1, wherein k is
 1. 176. Thecompound of claim 175, wherein the compound is:


177. The compound of claim 1, wherein k is
 2. 178. The compound of claim177, wherein the compound is: